Quinoline-substituted compound

ABSTRACT

An object to be achieved by the present invention is to provide a novel compound having EGFR inhibitory effects and cell growth inhibitory effects, as well as a medication useful for the prevention and/or treatment of cancer based on the EGFR inhibitory effects. 
     The present invention provides a compound represented by Formula (I) below, or a salt thereof.

CROSS REFERENCE TO RELATED APPLICATIONS

The present application is a divisional of U.S. Ser. No. 14/911,093filed Feb. 9, 2016, which is a U.S.C. 371 National Entry Applicationfrom PCT/JP2014/071951 filed on Aug. 22, 2014 which claims priority tothe specification of Japan Patent Application No. 2013-172746 (theentire disclosure is incorporated in the present specification byreference) filed on Aug. 22, 2013.

TECHNICAL FIELD

The present invention relates to quinoline-substituted compounds havingan inhibitory action against Epidermal Growth Factor Receptor (EGFR),and pharmaceutical compositions containing those as an activeingredient.

BACKGROUND ART

EGFR is a receptor-type tyrosine kinase, exerts its physiologicalfunction in normal tissue when being bound to Epidermal Growth Factor(EGF), which is a ligand, and contributes to growth, apoptosisinhibition, etc., in epithelial tissues (Non-Patent Document (NPD) 1).

In addition, EGFR is one of the oncogenes, and amplification of the EGFRgene and high expression or mutation of its protein are seen in variouscancer types such as head and neck cancer, breast cancer, colorectalcancer, esophagus cancer, pancreatic cancer, lung cancer, ovariancancer, renal cancer, bladder cancer, skin cancer, and brain tumor(Non-Patent Document (NPD) 2). In Japan and western countries,approximately 170 to 375 in every 100,000 people perish due to cancerevery year, and cancer ranks high as a cause of death (Non-PatentDocument (NPD) 3). Among these, the death toll due to lung cancerreaches approximately 1,400,000 per year worldwide, and since non-smallcell lung cancer accounts for 80% or more of lung cancers, there hasbeen a desire for the development of an effective therapy for the same(Non-patent Document (NPD) 4).

In recent years, responsible genes for these cancers are beingidentified, and a mutation in the EGFR gene is also one of them andresults in an active mutated EGFR protein. An active mutated EGFRprotein is, for example, a deletion of amino acid at positions 746-750(EGFR (d746-750)), a mutation of amino acid at position 858 from leucineto arginine (EGFR (L858R)), or the like. Such mutations are reported,for example, in 20-40% of non-small cell lung cancer cases in Japan, andin 10-15% of non-small cell lung cancer cases in western countries.Since non-small cell lung cancer having these mutations is highlysusceptible against gefitinib (product name: Iressa (Registeredtrademark)) and erlotinib (product name: Tarceva (Registered trademark))which are chemical agents (EGFR inhibitors) that inhibit the kinaseactivity of EGFR, these chemical agents are used as therapeutic agentsin Japan and western countries. However, the cancer acquires resistanceagainst gefitinib and erlotinib after 6 to 12 months from the beginningof use and therapeutic effect becomes weak. Therefore, this acquiredresistance has been a serious problem for treating non-small cell lungcancer having a highly-susceptible mutated EGFR. It has been revealedthat approximately 50% of the acquired resistance is due to emergence ofa resistant mutated EGFR protein (EGFR (d746-750/T790M) or EGFR(T790M/L858R)) having a second mutation in the EGFR gene resulting inamino acid at position 790 to change from threonine to methionine. Ithas been an important task to develop a therapeutic agent that iseffective against non-small cell lung cancer having this drug resistantmutated EGFR (Non-patent Document (NPD) 5).

On the other hand, skin abnormality and alimentary canal disorder arereported as common side effects of the EGFR inhibitors of gefitinib anderlotinib, which are clinically used as therapeutic agents at present,and of EGFR inhibitors such as BIBW2992 etc., which are under clinicaltrial. It is widely thought that these side effects are caused by theEGFR inhibitors inhibiting the activity of not only a mutated EGFRexpressed in non-small cell lung cancer, but also the activity of thewild-type EGFR (EGFR (WT)) expressed in the skin or alimentary canal(Non-Patent Document (NPD) 1). From a standpoint of side effectreduction, it is considered to be preferable to have a weak inhibitoryactivity against EGFR (WT) in normal tissues.

Thus, there is expectation of possibly suppressing growth of non-smallcell lung cancer cells having a drug resistant mutated EGFR throughadministration of a chemical agent having weaker inhibitory activityagainst the wild-type EGFR when compared to inhibitory activity againstthe drug resistant mutated EGFR whose amino acid at position 790 hasmutated to methionine, at an administration dose where the side effectto the skin or alimentary canal does not appear strongly. This ispredicted to contribute to treating the cancer, and prolonging life andimproving QOL of patients. In addition, if the chemical agent has weakinhibitory activity against the wild-type EGFR but has strong ininhibitory activity not only against drug resistant mutated EGFR butalso against highly-susceptible mutated EGFRs such as the EGFR(d746-750) and the EGFR (L858R) etc., which are highly susceptibleagainst gefitinib and erlotinib; there is expectation of possiblysuppressing growth of non-small cell lung cancer cells expressing ahighly-susceptible mutated EGFR or a drug resistant mutated EGFR at anadministration dose where the side effect to the skin or alimentarycanal does not appear strongly, or expectation of possibly reducing thefrequency of drug resistant mutated EGFR that emerges, as acquiredresistance, from non-small cell lung cancer expressing ahighly-susceptible mutated EGFR. This is predicted to contribute totreating the cancer, and prolonging life and improving QOL of patients.Furthermore, since expressions of highly-susceptible mutated EGFR anddrug resistant mutated EGFR can be used in the actual scene of therapyas indices for stratification to enable selection of patients, theycontribute greatly from an ethical viewpoint.

As a compound having a structure analogous to a compound according topresent invention,N-(3-(4-amino-6,7,8,9-tetrahydropyrimido[5,4-b]indolizin-5-yl)phenyl)benzamidederivative is known (Patent Document (PTD) 1). Although Patent Document1 describes using the amide compound for treating diseases characterizedby B-RAF kinase, the document does not disclose specific tests andresults therefrom corroborating a kinase inhibiting activity, and suchactivity is not confirmed.

CITATION LIST Patent Documents

PTD 1: International Publication No. W02006/102079 pamphlet

Non-Patent Documents

NPD 1: Nature Rev. Cancer, vol. 6, pp 803-811 (2006)

NPD 2: J. Clin. Oncol., vol. 19, 32s-40s (2001)

NPD 3: Ministry of Internal Affairs and Communications Statistics Bureauhomepage/statistical data/world statistics “World Statistics 2011”

NPD 4: Lung Cancer, vol. 69, pp 1-12 (2010)

NPD 5: Nature Rev. Cancer, vol. 10, pp 760-774 (2010)

SUMMARY OF INVENTION Technical Problem

As described above, EGFR inhibitors, although expected to be effectivein cancer therapy, are currently not clinically effective enough.

Therefore, an object of the present invention is to provide a novelcompound that strongly inhibits EGFR or a salt thereof. A further objectof the present invention is to provide: a novel compound that inhibitsmutated EGFR, for example, EGFR (d746-750), EGFR (L858R), EGFR(d746-750/T790M), and EGFR (T790M/L858R), but that does not inhibit EGFR(WT); or a salt thereof.

Solution to Problem

The present inventors have conducted thorough research in order toachieve the above-described object. As a result, they have found that agroup of quinoline-substituted compounds of the present invention haveexcellent inhibitory activity against EGFR and have cancer-cell-growthinhibitory action, and are useful as medication for treating cancer. Thepresent inventors thereby achieved the present invention.

Thus, the present invention provides the following items.

-   Item 1. A compound represented by Formula (I) below or a salt    thereof:

wherein the group:

is (1) a group represented by Formula A1:

(in Formula A1, B is a group represented by:

[R₁ is a hydrogen atom or a C1-C6 alkyl group; and R₂ is a grouprepresented by:

wherein R₃, R₄, and R₅ are the same or different, and each represents ahydrogen atom, a halogen atom, a C1-C6 alkyl group, a C6-C12 aryl group,a C4-C9 heteroaryl group, an aminomethyl group that may be substitutedwith a C1-C6 alkyl group, or a 1-piperidinomethyl group,

or a group represented by:

wherein R₆ represents a hydrogen atom or a C1-C6 alkyl group],

R₇ and R₈ are the same or different, and each represents a hydrogen atomor a C1-C6 alkyl group; m is 0 or 1; and n is 1 or 2);

(2) a group represented by Formula A2:

(in Formula A2, B and n are as defined in Formula A1; and R₉ is ahydrogen atom or a C1-C6 alkyl group); or

(3) a group represented Formula A3:

(in Formula A3, B, m, and n are as defined in Formula A1; and R₁₀ is aC1-C6 alkyl group).

-   Item 2. The compound or a salt thereof according to Item 1, wherein    R₂ is a group represented by:

wherein R₃, R₄, and R₅ are the same or different, and each represents ahydrogen atom, a halogen atom, a C1-C6 alkyl group, an aminomethyl groupthat may be substituted with a C1-C6 alkyl group, or a1-piperidinomethyl group,

or a group represented by:

wherein R₆ represents a hydrogen atom or a C1-C6 alkyl group.

-   Item 3. The compound or a salt thereof according to Item 1 or 2,    wherein R₂ is a group represented by:

wherein R₃, R₄, and R₅ are the same or different, and each represents ahydrogen atom, a halogen atom, an aminomethyl group that may besubstituted with a methyl group, or a 1-piperidinomethyl group.

-   Item 4. The compound or a salt thereof according to any one of Items    1 to 3, wherein the group:

is (1) a group represented by Formula A1:

(in Formula A1, B is a group represented by:

wherein R₁ is a hydrogen atom or a C1-C6 alkyl group; and R₂ is a grouprepresented by:

wherein R₃, R₄, and R₅ are the same or different, and each represents ahydrogen atom or a halogen atom,

R₇ and R₈ are the same or different, and each represents a hydrogen atomor a C1-C6 alkyl group; m is 0 or 1; and n is 1); or

(2) a group represented by Formula A2:

(in Formula A2, B and n are as defined in Formula A1; and R₉ is ahydrogen atom or a C1-C6 alkyl group).

-   Item 5. The compound or a salt thereof according to any one of Items    1 to 4, wherein the group:

is (1) a group represented by Formula A1:

(in Formula A1, B is a group represented by:

wherein R₁ is a hydrogen atom; and R₂ is a group represented by:

wherein R₃, R₄, and R₅ each represents a hydrogen atom,

R₇ and R₈ each represents a hydrogen atom; m is 0; and n is 1); or

(2) a group represented by Formula A2:

(in Formula A2, B and n are as defined in Formula A1; and R₉ representsa C1-C6 alkyl group).

-   Item 6. The compound or a salt thereof according to any one of Items    1 to 5, wherein the compound is selected from the following group of    compounds:-   (S)—N-(4-amino-6-methylene-5-(quinolin-3-yl)-7,8-dihydro-6H-pyrimido[5,4-b]pyrrolizin-7-yl)acrylamide    (Compound 1);-   (S)—N-(4-amino-6-methylene-5-(quinolin-3-yl)-7,8-dihydro-6H-pyrimido[5,4-b]pyrrolizin-7-yl)methacrylamide    (Compound 2);-   (S)—N-(4-amino-6-methylene-5-(quinolin-3-yl)-7,8-dihydro-6H-pyrimido[5,4-b]pyrrolizin-7-yl)but-2-enamide    (mixture of E and Z) (Compound 3);-   (S,E)-N-(4-amino-6-methylene-5-(quinolin-3-yl)-7,8-dihydro-6H-pyrimido[5,4-b]pyrrolizin-7-yl)-4-(dimethylamino)but-2-enamide    (Compound 4);-   (S,E)-N-(4-amino-6-methylene-5-(quinolin-3-yl)-7,8-dihydro-6H-pyrimido[5,4-b]pyrrolizin-7-yl)-3-chloroacrylamide    (Compound 5);-   (S,Z)—N-(4-amino-6-methylene-5-(quinolin-3-yl)-7,8-dihydro-6H-pyrimido[5,4-b]pyrrolizin-7-yl)-3-chloroacrylamide    (Compound 6);-   (S,E)-N-(4-amino-6-methylene-5-(quinolin-3-yl)-7,8-dihydro-6H-pyrimido[5,4-b]pyrrolizin-7-yl)-4-(piperidin-1-yl)but-2-enamide    (Compound 7);-   (S)—N-(4-amino-6-methylene-5-(quinolin-3-yl)-7,8-dihydro-6H-pyrimido[5,4-b]pyrrolizin-7-yl)propiolamide    (Compound 8);-   (S)—N-(4-amino-6-methylene-5-(quinolin-3-yl)-7,8-dihydro-6H-pyrimido[5,4-b]pyrrolizin-7-yl)but-2-ynamide    (Compound 9);-   (S,E)-N-(4-amino-6-methylene-5-(quinolin-3-yl)-7,8-dihydro-6H-pyrimido[5,4-b]pyrrolizin-7-yl)-4-(diethylamino)but-2-enamide    (Compound 10);-   (S,E)-N-(4-amino-6-methylene-5-(quinolin-3-yl)-7,8-dihydro-6H-pyrimido[5,4-b]pyrrolizin-7-yl)-4-(ethyl(methyl)amino)but-2-enamide    (Compound 11);-   (S,E)-N-(4-amino-6-methylene-5-(quinolin-3-yl)-7,8-dihydro-6H-pyrimido[5,4-b]pyrrolizin-7-yl)-4-(isopropyl(methyl)amino)but-2-enamide    (Compound 12);-   (R)—N-(4-amino-6-methylene-5-(quinolin-3-yl)-6,7,8,9-tetrahydropyrimido[5,4-b]indolizin-7-yl)acrylamide    (Compound 13);-   (S)—N-(4-amino-6-methyl-5-(quinolin-3-yl)-8,9-dihydropyrimido[5,4-b]indolizin-8-yl)acrylamide    (Compound 14);-   (S)—N-(4-amino-6-methylene-5-(quinolin-3-yl)-6,7,8,9-tetrahydropyrimido[5,4-b]indolizin-8-yl)acrylamide    (Compound 15);-   (R)—N-(4-amino-6-methyl-5-(quinolin-3-yl)-8,9-dihydropyrimido[5,4-b]indolizin-8-yl)acrylamide    (Compound 16);-   (R)—N-(4-amino-6-methylene-5-(quinolin-3-yl)-6,7,8,9-tetrahydropyrimido[5,4-b]indolizin-8-yl)acrylamide    (Compound 17);-   (S)—N-(4-amino-6-methylene-5-(quinolin-3-yl)-7,8,9,10-tetrahydro-6H-pyrimido[5′,4′:4,5]pyrrolo[1,2-a]azepin-8-yl)acrylamide    (Compound 18);-   (S,E)-N-(4-amino-6-ethylidene-5-(quinolin-3-yl)-6,7,8,9-tetrahydropyrimido[5,4-b]indolizin-8-yl)acrylamide    (Compound 19);-   (S)—N-(4-amino-6-isopropyl-5-(quinolin-3-yl)-8,9-dihydropyrimido[5,4-b]indolizin-8-yl)acrylamide    (Compound 20A);-   (S)—N-(4-amino-6-(propan-2-ylidene)-5-(quinolin-3-yl)-6,7,8,9-tetrahydropyrimido[5,4-b]indolizin-8-yl)acrylamide    (Compound 20B);-   (R)—N-(4-amino-6-methylene-5-(quinolin-3-yl)-7,8-dihydro-6H-pyrimido[5,4-b]pyrrolizin-7-yl)-N-methylacrylamide    (Compound 21);-   (R)—N-(4-amino-6-methylene-5-(quinolin-3-yl)-6,7,8,9-tetrahydropyrimido[5,4-b]indolizin-8-yl)-N-methylacrylamide    (Compound 22);-   (R)—N-(4-amino-6-methyl-5-(quinolin-3-yl)-8,9-dihydropyrimido[5,4-b]indolizin-8-yl)-N-methylacrylamide    (Compound 23);-   N-((7S)-4-amino-6-methyl-5-(quinolin-3-yl)-7,8-dihydro-6H-pyrimido[5,4-b]pyrrolizin-7-yl)acrylamide    (Compound 24);-   (R)—N-(4-amino-6-methylene-5-(quinolin-3-yl)-7,8-dihydro-6H-pyrimido[5,4-b]pyrrolizin-7-yl)acrylamide    (Compound 25);-   (S)—N-(4-amino-6-methylene-5-(quinolin-3-yl)-7,8-dihydro-6H-pyrimido[5,4-b]pyrrolizin-7-yl)-N-methylacrylamide    (Compound 26);-   (S)—N-(4-amino-5-(quinolin-3-yl)-8,9-dihydropyrimido[5,4-b]indolizin-8-yl)acrylamide    (Compound 27);-   (R)—N-(4-amino-5-(quinolin-3-yl)-9,10-dihydro-8H-pyrimido[5′,    4′:4,5]pyrrolo[1,2-a]azepin-8-yl)acrylamide (Compound 28);-   N-((6R*,8S)-4-amino-6-methyl-5-(quinolin-3-yl)-6,7,8,9-tetrahydropyrimido[5,4-b]indolizin-8-yl)acrylamide    (Compound 29A); and-   N-((6S*,8S)-4-amino-6-methyl-5-(quinolin-3-yl)-6,7,8,9-tetrahydropyrimido[5,4-b]indolizin-8-yl)acrylamide    (Compound 29B).-   Item 7. The compound or a salt thereof according to any one of Items    1 to 5, wherein the compound is selected from the following group of    compounds:-   (S)—N-(4-amino-6-methylene-5-(quinolin-3-yl)-7,8-dihydro-6H-pyrimido[5,4-b]pyrrolizin-7-yl)acrylamide    (Compound 1);-   (S,E)-N-(4-amino-6-methylene-5-(quinolin-3-yl)-7,8-dihydro-6H-pyrimido[5,4-b]pyrrolizin-7-yl)-3-chloroacrylamide    (Compound 5);-   (S,Z)—N-(4-amino-6-methylene-5-(quinolin-3-yl)-7,8-dihydro-6H-pyrimido[5,4-b]pyrrolizin-7-yl)-3-chloroacrylamide    (Compound 6);-   (S)—N-(4-amino-6-methyl-5-(quinolin-3-yl)-8,9-dihydropyrimido[5,4-b]indolizin-8-yl)acrylamide    (Compound 14);-   (S)—N-(4-amino-6-methylene-5-(quinolin-3-yl)-6,7,8,9-tetrahydropyrimido[5,4-b]indolizin-8-yl)acrylamide    (Compound 15);-   (S,E)-N-(4-amino-6-ethylidene-5-(quinolin-3-yl)-6,7,8,9-tetrahydropyrimido[5,4-b]indolizin-8-yl)acrylamide    (Compound 19);-   (R)—N-(4-amino-6-methylene-5-(quinolin-3-yl)-7,8-dihydro-6H-pyrimido[5,4-b]pyrrolizin-7-yl)-N-methylacrylamide    (Compound 21);-   (R)—N-(4-amino-6-methylene-5-(quinolin-3-yl)-6,7,8,9-tetrahydropyrimido[5,4-b]indolizin-8-yl)-N-methylacrylamide    (Compound 22);-   (R)—N-(4-amino-6-methyl-5-(quinolin-3-yl)-8,9-dihydropyrimido[5,4-b]indolizin-8-yl)-N-methylacrylamide    (Compound 23); and-   (R)—N-(4-amino-5-(quinolin-3-yl)-9,10-dihydro-8H-pyrimido[5′,4′:4,5]pyrrolo[1,2-a]azepin-8-yl)acrylamide    (Compound 28).-   Item 8.    (S)—N-(4-amino-6-methylene-5-(quinolin-3-yl)-7,8-dihydro-6H-pyrimido[5,4-b]pyrrolizin-7-yl)acrylamide    (Compound 1) or a salt thereof.-   Item 9.    (S)—N-(4-amino-6-methyl-5-(quinolin-3-yl)-8,9-dihydropyrimido[5,4-b]indolizin-8-yl)acrylamide    (Compound 14) or a salt thereof.-   Item 10. An EGFR inhibitor comprising the compound or a salt thereof    according to any one of Items 1 to 9, as an active ingredient.-   Item 11. A pharmaceutical composition comprising the compound or a    salt thereof according to any one of Items 1 to 9.-   Item 12. An antitumor agent comprising the compound or a salt    thereof according to any one of Items 1 to 9, as an active    ingredient.-   Item 13. A method for treating or preventing cancer, the method    comprising the step of administering, to a mammal, the compound or a    salt thereof according to any one of Items 1 to 9 at a dose    effective for treating or preventing cancer.-   Item 14. Use of the compound or a salt thereof according to any one    of Items 1 to 9 in the manufacture of an antitumor agent.-   Item 15. The compound or a salt thereof according to any one of    Items 1 to 9 for use in the prevention or treatment of cancer.

Advantageous Effects of Invention

According to the present invention, a novel compound represented byFormula (I) above or a salt thereof useful as an EGFR inhibitor isprovided.

It has been clarified that the compound of the present invention or asalt thereof has excellent EGFR inhibition activity and a cancer cellstrain growth inhibitory effect. In addition, the compound of thepresent invention or a salt thereof has advantageously few side effectsas a result of excellent selectivity against EGFRs. Therefore, thecompound of the present invention or a salt thereof is useful as anagent for preventing and/or treating cancer.

DESCRIPTION OF EMBODIMENTS

The compound represented by Formula (I) according to the presentinvention is a quinoline-substituted compound having a quinolinestructure and an α, β-unsaturated amide structure, and is a novelcompound nowhere disclosed in any of the above-mentioned prior artdocuments etc.

Specifically, the compound specifically disclosed in PTD 1 is anN-(3-(4-amino-6,7,8,9-tetrahydropyrimido[5,4-b]indolizin-5-yl)phenyl)benzamidederivative. The compound of the present invention is different from thecompound disclosed in PTD 1 in that the compound of the presentinvention has a quinoline structure and an α, β-unsaturated amidestructure.

In the specification of the present invention, examples of the “halogenatom” include fluorine, chlorine, bromine, and iodine.

In the specification of the present invention, the term “C1-C6 alkylgroup” refers to a linear or branched alkyl group having 1 to 6 carbonatoms. Specific examples thereof include methyl, ethyl, n-propyl,isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, pentyl, and hexyl.

In the specification of the present invention, the term “C6-C12 arylgroup” refers to an aryl group having 6 to 12 carbon atoms. Specificexamples thereof include phenyl, naphthyl, and biphenyl.

In the specification of the present invention, the term “C4-C9heteroaryl group” refers to a monocyclic or bicyclic C4-C9 heteroarylgroup containing 1 to 3 same or different heteroatoms selected fromnitrogen, oxygen, and sulfur atoms; and is preferably a monocyclic orbicyclic C4-C9 heteroaryl group containing 1 to 3 nitrogen atoms.Specific examples thereof include thienyl, furyl, pyrrolyl, triazolyl,imidazolyl, pyrazolyl, isothiazolyl, isoxazolyl, pyridyl, pyrazinyl,pyrimidinyl, pyridazinyl, isobenzofuryl, indolizinyl, isoindolyl,indolyl, indazolyl, quinolyl, isoquinolyl, phthalazinyl, andnaphthyridinyl.

In the specification of the present invention, the term “aminomethylgroup that may be substituted with a C1-C6 alkyl group” refers to anaminomethyl group in which at least one hydrogen atom of the aminomoiety may be substituted with a linear or branched alkyl group having 1to 6 carbon atoms. Specific examples include aminomethyl,N-methylaminomethyl, N,N-dimethylaminomethyl, N-ethylaminomethyl,N,N-diethylaminomethyl, N-methylN-ethylaminomethyl,N-methylN-isopropylaminomethyl, N-propylaminomethyl, N-butylaminomethyl,N-pentylaminomethyl, and N-hexylaminomethyl.

In Formula (I) above, the moiety represented by:

is (1) a group represented by Formula A1:

(in Formula A1, B, m, n, R₇, and R₈ are as defined above);

(2) a group represented by Formula A2:

(in Formula A2, B, n, and R₉ are as defined above); or

(3) a group represented by Formula A3:

(in Formula A3, B, m, n, R₇, and R₈ are as defined above),

and is preferably (1) the group represented by Formula A1:

(in the formula, B, m, n, R₇, and R₈ are as defined above), or (2) thegroup represented by Formula A2:

(in the formula, B, n and R₉ are as defined above)

m in Formula (I) is preferably 0.

n in Formula (I) is preferably 1.

R₁ in Formula (I) is preferably a hydrogen atom.

R₂ in Formula (I) is preferably a group represented by:

(wherein R₃, R₄, and R₅ are as defined above).

R₃, R₄, and R₅ in Formula (I) are preferably the same or different, andeach represents a hydrogen atom, a halogen atom, a C1-C6 alkyl group, anaminomethyl group that may be substituted with a C1-C6 alkyl group, or a1-piperidinomethyl group; more preferably, R₃, R₄, and R₅ are the sameor different, and each represents a hydrogen atom, a halogen atom, anaminomethyl group that may be substituted with a methyl group, or a1-piperidinomethyl group; further preferably, R₃, R₄, and R₅ are thesame or different, and each represents a hydrogen atom or a halogenatom, particular preferably a hydrogen atom.

R₇ and R₈ in Formula (I) are preferably the same or different, and eachrepresents a hydrogen atom or a C1-C6 alkyl group (preferably a C1-C3alkyl group, more preferably a methyl group); further preferably, atleast one of R₇ and R₈ is a hydrogen atom; particular preferably, bothof R₇ and R₈ are hydrogen atoms.

R₉ in Formula (I) is preferably a C1-C6 alkyl group, more preferably aC1-C3 alkyl group, further preferably a methyl group.

The compound of the present invention or a salt thereof preferably has astrong enzyme inhibitory activity against EGFR (T790M/L858R); morepreferably, the concentration of the compound by which 50% of the enzymecan be inhibited is 2 nM or less. Further, the compound of the presentinvention or a salt thereof preferably has a strong enzyme inhibitoryactivity against EGFR (d746-750/T790M); the 50% inhibitory concentrationof the compound is also preferably 2 nM or less. Further, the compoundof the present invention or a salt thereof preferably has a strong cellgrowth inhibitory effect against tumor cells with EGFR (T790M/L858R);the compound more preferably has a 50% inhibitory concentration of 200nM or less, further preferably 100 nM or less, and particular preferably40 nM or less.

Subsequently, the production method of the compound of the presentinvention is explained.

The compound (I) of the present invention may be produced, for example,through the following production method, the methods described in theExamples, and the like. However, the production method of the compoundof the present invention is not limited to these reaction examples.

Production Method 1

wherein P₁ is a protecting group of an amino group; L₁ is a leavinggroup; and R₂, R₇, R₈, m, and n are as defined above.

-   Step a

In this step, the compound represented by Formula (IV) is producedthrough a Mitsunobu reaction using the compounds represented by Formulas(II) and (III).

Examples of the leaving group represented by L₁ in the compoundrepresented by Formula (II) include a bromine or iodine atom. Thecompound represented by Formula (II) may be a commercially availableproduct, or can be produced by a known method. Examples of theprotecting group of an amino group represented by P₁ in Formula (III)include a tert-butoxycarbonyl group and a benzoyl group. The compoundrepresented by Formula (III) may be a commercially available product, orcan be produced by a known method. The compound represented by Formula(III) can be used in an amount of 1 to 10 moles, and preferably 1 to 5moles, per mole of the compound represented by Formula (II).

The Mitsunobu reaction may be performed according to a known method (forexample, the method disclosed in Synthesis, p. 1, 1981) or a similarmethod.

Examples of azodicarboxylic acid esters include diethyl azodicarboxylateand diisopropyl azodicarboxylate. Such an azodicarboxylic acid ester canbe used in an amount of 1 to 10 moles, and preferably 1 to 5 moles, permole of the compound represented by Formula (II).

As the phosphine compound, triphenylphosphine, tributylphosphine, or thelike can be used. The phosphine compound can be used in an amount of 1to 10 moles, and preferably 1 to 5 moles, per mole of the compoundrepresented by Formula (II).

As a solvent, tetrahydrofuran, 1,2-dimethoxyethane, 1,4-dioxane,toluene, N,N-dimethylformamide, N,N-dimethylacetamide, dimethylsulfoxide, N-methylpyrrolidin-2-one, and the like can be used alone, oras a mixture. The reaction time is 0.1 to 100 hours, and preferably 0.1to 24 hours. The reaction temperature is 0° C. to the boilingtemperature of the solvent, and preferably 0° C. to 100° C.

The thus-obtained compound represented by Formula (IV) can be subjectedto the subsequent step after or without isolation or purification byknown isolation and purification means, such as concentration, vacuumconcentration, crystallization, solvent extraction, reprecipitation, andchromatography.

-   Step b

In this step, the compound represented by Formula (IV) is reacted withammonia or a salt thereof to produce the compound represented by Formula(V).

The amount of ammonia or a salt thereof used in this step is typicallyan equimolar to excessive molar amount per mole of the compoundrepresented by Formula (IV).

Any reaction solvent that does not adversely affect the reaction can beused. Examples of usable reaction solvents include water, methanol,ethanol, isopropanol, tert-butyl alcohol, tetrahydrofuran,1,2-dimethoxyethane, 1,4-dioxane, N,N-dimethylformamide,N-methylpyrrolidin-2-one, dimethyl sulfoxide, and mixed solventsthereof.

The reaction temperature is typically 0° C. to 200° C., preferably fromroom temperature to 150° C. The reaction time is typically 5 minutes to7 days, and preferably 30 minutes to 24 hours.

The thus-obtained compound represented by Formula (V) can be subjectedto the subsequent step after or without isolation and purification byknown isolation and purification means, such as concentration, vacuumconcentration, crystallization, solvent extraction, reprecipitation, andchromatography.

-   Step c

In this step, the compound represented by Formula (V) is subjected to acoupling reaction with 3-quinolineboronic acid or 3-quinolineboronicacid ester to produce the compound represented by Formula (VI).

This step can be performed according to a generally known method (forexample, Chemical Reviews, Vol. 95, p. 2457, 1995). This step can beperformed in the presence of a transition metal catalyst and a base in asolvent that does not adversely affect the reaction.

The amount of 3-quinolineboronic acid or 3-quinolineboronic acid esterused may be 1 to 10 moles, and preferably 1 to 3 moles, per mole of thecompound represented by Formula (V).

Examples of transition metal catalysts include palladium catalysts(e.g., palladium acetate, palladium chloride,tetrakis(triphenylphosphine)palladium,1,1′-bis(diphenylphosphino)ferrocene-palladium (II) dichloride, andtris(dibenzylideneacetone)dipalladium (0)), nickel catalysts (e.g.,nickel chloride), and the like. If necessary, a ligand (e.g.,triphenylphosphine, tri-tert-butylphosphine, or2-dicyclohexylphosphino-2′,4′,6′-triisopropylbiphenyl) can be added, anda metal oxide (such as copper oxide or silver oxide) can be used as aco-catalyst. The amount of the transition metal catalyst used may varydepending on the type of catalyst. The transition metal catalyst istypically used in an amount of 0.0001 to 1 mole, and preferably 0.01 to0.5 moles, per mole of the compound represented by Formula (V). Theamount of the ligand used is typically 0.0001 to 4 moles, and preferably0.01 to 2 moles, per mole of the compound represented by Formula (V).The amount of the co-catalyst used is typically 0.0001 to 4 moles, andpreferably 0.01 to 2 moles, per mole of the compound represented byFormula (V).

Examples of usable bases include organic amines (e.g., trimethylamine,triethylamine, diisopropylethylamine, N-methylmorpholine,1,8-diazabicyclo[5,4,0]undec-7-ene, pyridine, and N,N-dimethylaniline),alkali metal salts (e.g., sodium hydrogen carbonate, potassium hydrogencarbonate, sodium carbonate, potassium carbonate, cesium carbonate,sodium phosphate, potassium phosphate, sodium hydroxide, and potassiumhydroxide), metal hydrides (e.g., potassium hydride and sodium hydride),alkali metal alkoxides (e.g., sodium methoxide, sodium ethoxide, sodiumtert-butoxide, and potassium tert-butoxide), alkali metal disilazides(e.g., lithium disilazide, sodium disilazide, and potassium disilazide),and the like. Among them, alkali metal salts such as sodium carbonate,potassium carbonate, cesium carbonate, sodium phosphate, and potassiumphosphate; alkali metal alkoxides such as sodium tert-butoxide andpotassium tert-butoxide; and organic amines such as triethylamine anddiisopropylethylamine are preferable. The amount of the base used istypically 0.1 to 10 moles, and preferably 1 to 5 moles, per mole of thecompound represented by Formula (V).

Any solvent that does not adversely affect the reaction can be used.Examples of usable solvent include hydrocarbons (e.g., benzene, toluene,and xylene), halogenated hydrocarbons (e.g., chloroform and1,2-dichloroethane), nitriles (e.g., acetonitrile), ethers (e.g.,1,2-dimethoxyethane, tetrahydrofuran, and 1,4-dioxane), alcohols (e.g.,methanol and ethanol), aprotic polar solvents (e.g.,N,N-dimethylformamide, dimethyl sulfoxide, and hexamethyl phosphorylamide), water, and mixed solvents thereof. The reaction time is 0.1 to100 hours, and preferably 0.5 to 24 hours. The reaction temperature is0° C. to the boiling temperature of the solvent, and preferably 20 to150° C.

The thus-obtained compound represented by Formula (VI) can be subjectedto the subsequent step after or without isolation and purification byknown isolation and purification means, such as concentration, vacuumconcentration, crystallization, solvent extraction, reprecipitation, andchromatography.

-   Step d

In this step, the compound represented by Formula (VI) is brominated bybeing reacted with N-bromosuccinimide to produce the compoundrepresented by Formula (VII).

The halogenation can be performed by the method disclosed in WO2006/102079, or by a similar method.

The amount of N-bromosuccinimide used in this step is 0.5 to 2.0 moles,and preferably 0.9 to 1.2 moles, per mole of the compound represented byFormula (VI).

Any reaction solvent that does not adversely affect the reaction can beused. For example, tetrahydrofuran, 1,4-dioxane, N,N-dimethylformamide,N-methylpyrrolidin-2-one, or a mixed solvent thereof can be preferablyused.

The reaction temperature is typically −20 to 50° C., and preferably 0°C. to room temperature. The reaction time is typically 1 minute to 2days, and preferably 5 minutes to 12 hours.

The thus-obtained compound represented by Formula (VI) can be subjectedto the subsequent step after or without isolation or purification byknown isolation and purification means, such as concentration, vacuumconcentration, crystallization, solvent extraction, reprecipitation, andchromatography.

-   Step e

This step subjects the compound represented by Formula (VII) to anintramolecular cyclization reaction to produce the compound representedby Formula (VIII).

This step can be performed according to a generally known method (forexample, the method disclosed in Chemical Reviews, Vol. 103, p. 2945,2003).

Examples of transition metal catalysts include bivalent palladiumcatalysts (e.g., palladium acetate, palladium chloride,1,1′-bis(diphenylphosphino)ferrocene-palladium(II)dichloride, etc.), andzero-valent palladium catalysts (e.g.,tetrakis(triphenylphosphine)palladium,tris(dibenzylideneacetone)dipalladium, etc.). If necessary, a ligand(e.g., triphenylphosphine, tri-tert-butylphosphine, etc.) can be added.The amount of the transition metal catalyst used may vary depending onthe type of catalyst. The transition metal catalyst is typically used inan amount of 0.0001 to 1 mole, and preferably 0.01 to 0.5 moles, permole of the compound represented by Formula (VII). The amount of theligand used is typically 0.0001 to 4 moles, and preferably 0.01 to 2moles, per mole of the compound represented by Formula (VII).

Examples of usable bases include inorganic bases such as sodium hydrogencarbonate, sodium carbonate, potassium carbonate, cesium carbonate,sodium hydroxide, potassium hydroxide, cesium hydroxide, sodium hydride,or potassium hydride. Such a base can be used in an amount of 1 to 100moles, and preferably 2 to 20 moles, per mole of the compoundrepresented by Formula (VII).

Any solvent that does not adversely affect the reaction can be used.Examples of usable solvents include hydrocarbons (e.g., benzene,toluene, and xylene), ethers (e.g., 1,2-dimethoxyethane,tetrahydrofuran, and 1,4-dioxane), aprotic polar solvents (e.g.,N,N-dimethylformamide, dimethyl sulfoxide, and hexamethyl phosphorylamide), water, and mixed solvents thereof. The reaction time is 0.1 to100 hours, and preferably 0.5 to 24 hours. The reaction temperature is-20° C. to the boiling temperature of the solvent, and preferably 0° C.to 150° C.

The thus-obtained compound represented by Formula (VIII) can besubjected to the subsequent step after or without isolation orpurification by known isolation and purification means, such asconcentration, vacuum concentration, crystallization, solventextraction, reprecipitation, and chromatography.

-   Step f

In this step, the protected amino group of the compound represented byFormula (VIII) is deprotected to produce the compound represented byFormula (IX).

The deprotection can be performed by a known method, such as the methoddescribed in Protective Groups in Organic Synthesis, T. W. Greene, JohnWiley & Sons (1981); or a similar method.

When a tert-butoxycarbonyl group is used as a protecting group,hydrochloric acid, sulfuric acid, methanesulfonic acid, trifluoroaceticacid, and the like are used as a deprotection reagent. The amount of thereagent used is preferably 1 to 100 moles per mole of the compound(VIII).

Any solvent that does not adversely affect the reaction can be used.Examples of usable solvents include water, methanol, ethanol, methylenechloride, chloroform, and the like, and mixed solvents thereof. Thereaction time is 0.1 to 100 hours, and preferably 0.5 to 24 hours. Thereaction temperature is 0° C. to the boiling temperature of the solvent,and is preferably from 0 to 50° C.

The thus-obtained compound represented by Formula (IX) can be subjectedto the subsequent step after or without isolation and purification byknown isolation and purification means, such as concentration, vacuumconcentration, crystallization, solvent extraction, reprecipitation, andchromatography.

-   Step g

In this step, the compound represented by Formula (IX) is amidated withan α, β-unsaturated carboxylic acid, or α, (β-unsaturated acid chlorideor bromide to produce the compound represented by Formula (I-1) of thepresent invention.

When a carboxylic acid is used as an amidation reagent, the carboxylicacid can be used in an amount of 0.5 to 10 moles, preferably 1 to 3moles, per mole of the compound represented by Formula (IX), in thepresence of a suitable condensing agent. The carboxylic acid may be acommercially available product, or can be produced according to a knownmethod.

Any reaction solvent that does not adversely affect the reaction can beused. Examples of usable solvent include toluene, benzene, methylenechloride, chloroform, tetrahydrofuran, 1,4-dioxane,N,N-dimethylformamide, dimethylacetamide, N-methylpyrrolidin-2-one,dimethyl sulfoxide, and mixed solvents thereof. The reaction temperatureis typically −78 to 200° C., and preferably 0 to 50° C. The reactiontime is typically 5 minutes to 3 days, and preferably 5 minutes to 10hours.

Examples of condensation agents include diphenylphosphoryl azide,N,N′-dicyclohexylcarbodiimide,benzotriazol-1-yloxy-trisdimethylaminophosphonium salts,4-(4,6-dimethoxy-1,3,5-triazin-2-yl)-4-methylmorpholinium chloride,1-ethyl-3-(3-dimethylaminopropyl)carbodiimide, a combination of1-ethyl-3-(3-dimethylaminopropyl)carbodiimide and1-hydroxybenzotriazole, 2-chloro-1,3-dimethylimidazolinium chloride,O-(7-azabenzotriazo-1-yl)-N,N,N′,N′-tetramethylhexauroniumhexafluorophosphate, and the like.

If necessary, a base can be optionally added for the reaction. Examplesof usable bases include organic bases such as triethylamine,diisopropylethylamine, pyridine, lutidine, collidine,4-(N,N-dimethylamino)pyridine, potassium tert-butyrate, sodiumtert-butyrate, sodium methoxide, sodium ethoxide, lithiumhexamethyldisilazide, sodium hexamethyldisilazide, potassiumhexamethyldisilazide, and butyl lithium; and inorganic bases such assodium hydrogen carbonate, sodium carbonate, potassium carbonate, cesiumcarbonate, sodium hydroxide, and sodium hydride. Such a base can beadded in an amount of 1 to 100 moles, and preferably 1 to 10 moles, permole of the compound represented by Formula (IX).

When an acid chloride or acid bromide is used as an amidation reagent,the acid halide is used in an amount of 0.5 to 5 moles, and preferably0.9 to 1.1 moles, per mole of the compound represented by Formula (IX).The acid halide may be a commercially available product, or can beproduced according to a known method.

Any reaction solvent that does not adversely affect the reaction can beused. Examples thereof include toluene, benzene, methylene chloride,chloroform, tetrahydrofuran, 1,4-dioxane, N,N-dimethylformamide,dimethylacetamide, N-methylpyrrolidin-2-one, acetonitrile, water, andmixed solvents thereof. The reaction temperature is typically −78 to200° C., preferably 0 to 50° C. The reaction time is typically 5 minutesto 3 days, and preferably 5 minutes to 10 hours.

If necessary, a base can be added for the reaction. Examples of usablebases include organic bases such as triethylamine,diisopropylethylamine, pyridine, lutidine, collidine,4-(N,N-dimethylamino)pyridine, potassium tert-butyrate, sodiumtert-butyrate, sodium methoxide, sodium ethoxide, lithiumhexamethyldisilazide, sodium hexamethyldisilazide, potassiumhexamethyldisilazide, and butyl lithium; and inorganic bases such assodium hydrogen carbonate, sodium carbonate, potassium carbonate, cesiumcarbonate, sodium hydroxide, and sodium hydride. Such a base can beadded in an amount of 1 to 100 moles, and preferably 1 to 20 moles, permole of the compound represented by Formula (IX). The thus-obtainedcompound represented by Formula (I-1) can be isolated and purified byknown isolation and purification means, such as concentration, vacuumconcentration, crystallization, solvent extraction, reprecipitation, andchromatography.

Production Method 2

wherein P₁ represents a protecting group of an amino group; and R₂, R₇,R₈, R₉, and n are as defined above.

-   Step h

In this step, the protected amino group of the compound represented byFormula (X) is deprotected to produce the compound represented byFormula (XI).

The deprotection can be performed by a known method, such as the methoddescribed in Protective Groups in Organic Synthesis, T. W. Greene, JohnWiley & Sons (1981); or a similar method.

When tert-butoxycarbonyl is used as a protecting group, hydrochloricacid, sulfuric acid, methanesulfonic acid, trifluoroacetic acid, or thelike can be used as a deprotection reagent. The reagent is preferablyused in an amount of 1 to 100 moles per mole of Compound (X).

Any solvent that does not adversely affect the reaction can be used.Examples of usable solvents include water, methanol, ethanol, methylenechloride, chloroform, and mixed solvents thereof. The reaction time is0.1 to 100 hours, and preferably 0.5 to 24 hours. The reactiontemperature is 0° C. to the boiling temperature of the solvent, and ispreferably from 50° C. to the boiling temperature of the solvent.

The thus-obtained compound represented by Formula (XXI) can be subjectedto the subsequent step after or without isolation and purification byknown isolation and purification means, such as concentration, vacuumconcentration, crystallization, solvent extraction, reprecipitation, andchromatography.

-   Step i

This step can be performed in the same manner as in Step g.

Production Method 3

wherein P₁ represents a protecting group of an amino group; L₁ and L₂represent a leaving group; and R₁, R₂, R₇, R₈, m, and n are as definedabove.

-   Step j

In this step, the compound represented by Formula (XIII) is producedthrough an alkylation reaction using the compounds represented byFormulas (IV) and (XII) in the presence of a base.

In the compound represented by Formula (XII), examples of the leavinggroup represented by L₂ include bromine atom, iodine atom,methanesulfonic acid ester, p-toluenesulfonic acid ester, and the like.The compound represented by Formula (XII) may be a commerciallyavailable product, or can be produced by a known method. The compoundrepresented by Formula (XII) can be used in an amount of 1 to 10 moles,and preferably 1 to 5 moles, per mole of the compound represented byFormula (IV). Examples of usable bases include organic amines (e.g.,trimethylamine, triethylamine, diisopropylethylamine,N-methylmorpholine, 1,8-diazabicyclo[5,4,0]undec-7-ene, pyridine, andN,N-dimethylaniline), alkali metal salts (e.g., sodium hydrogencarbonate, potassium hydrogen carbonate, sodium carbonate, potassiumcarbonate, cesium carbonate, sodium phosphate, potassium phosphate,sodium hydroxide, and potassium hydroxide), metal hydrides (e.g.,potassium hydride and sodium hydride), alkali metal alkoxides (e.g.,sodium methoxide, sodium ethoxide, sodium tert-butoxide, and potassiumtert-butoxide), alkali metal disilazides (e.g., lithium disilazide,sodium disilazide, and potassium disilazide), and the like. Among them,alkali metal salts such as sodium carbonate, potassium carbonate, cesiumcarbonate, sodium phosphate, and potassium phosphate; and metal hydridessuch as sodium hydride; alkali metal alkoxides such as sodiumtert-butoxide and potassium tert-butoxide are preferable. The amount ofthe base used is typically 0.1 to 10 moles, and preferably 1 to 5 moles,per mole of the compound represented by Formula (V).

As a solvent, tetrahydrofuran, 1,2-dimethoxyethane, 1,4-dioxane,N,N-dimethylformamide, N,N-dimethylacetamide, dimethyl sulfoxide,N-methylpyrrolidin-2-one, and the like can be used alone, or as amixture. The reaction time is 0.1 to 100 hours, and preferably 0.5 to 24hours. The reaction temperature is 0° C. to the boiling temperature ofthe solvent, and is preferably from 20° C. to 150° C.

The thus-obtained compound represented by Formula (XIII) can besubjected to the subsequent step after or without isolation orpurification by known isolation and purification means, such asconcentration, vacuum concentration, crystallization, solventextraction, reprecipitation, and chromatography.

-   Step k

This step can be performed in the same manner as in Step b.

-   Step l

This step can be performed in the same manner as in Step c.

-   Step m

This step can be performed in the same manner as in Step d.

-   Step n

This step can be performed in the same manner as in Step e.

-   Step o

This step can be performed in the same manner as in Step f.

-   Step p

This step can be performed in the same manner as in Step g.

Production Method 4

wherein P₁ represents a protecting group of an amino group; and R₁, R₂,R₇, R₈, R₉, and n are as defined above.

-   Step q

This step can be performed in the same manner as in Step h.

-   Step r

This step can be performed in the same manner as in Step g.

Production Method 5

wherein P₁ represents a protecting group of an amino group; and R₁, R₂,R₂, R₈, R₁₀, m, and n are as defined above.

-   Step s

In this step, the compound represented by Formula (XX) is producedthrough hydrogenation with respect to the compound represented byFormula (XVII) in the presence of a catalyst.

Examples of catalyst include palladium-carbon catalyst, palladiumhydroxide-carbon catalyst, and Raney nickel catalyst. The catalyst canbe used in an amount of 0.01 moles to an excessive amount, preferablyfrom 0.1 moles to 10 moles, per mole of the compound represented byFormula (XVII).

The hydrogenation can be performed at 1 atm to 100 atm, preferably 1 atmto 10 atm. As a solvent, methanol, ethanol, ethyl acetate,tetrahydrofuran, 1,2-dimethoxyethane, 1,4-dioxane,N,N-dimethylformamide, N,N-dimethylacetamide, N-methylpyrrolidin-2-one,water, and the like can be used alone, or as a mixture. The reactiontime is from 0.1 to 100 hours, preferably from 0.5 to 48 hours. Thereaction temperature ranges from room temperature to the boilingtemperature of the solvent, preferably from room temperature to 100° C.

The thus-obtained compound represented by Formula (XX) can be subjectedto the subsequent step after or without isolation or purification byknown isolation and purification means, such as concentration, vacuumconcentration, crystallization, solvent extraction, reprecipitation, andchromatography.

-   Step t

This step can be performed in the same manner as in Step f.

-   Step u

This step can be performed in the same manner as in Step g.

Production Method 6

-   wherein P₁ represents a protecting group of an amino group; and R₂    and n are as defined above.-   Step v

In this step, an organic borane reagent is allowed to act on thecompound represented by Formula (XXII) to prepare an alkyl boraneintermediate in the system, and then the compound represented by Formula(XXIII) is produced in the presence of a transition metal catalyst and abase.

This step can be performed according to a generally known method (forexample, WO 2006/102079).

Examples of organic borane reagents include9-BBN(9-borabicyclo[3.3.1]-nonane), 9-BBN(9-borabicyclo[3.3.1]-nonane)dimer, disiamylborane(bis(1,2-dimethylpropyl)borane),thexylborane(1,1,2-trimethylpropyl)borane), and the like. The organicborane reagent is preferably 9-BBN(9-borabicyclo[3.3.1]-nonane) or9-BBN(9-borabicyclo[3.3.1]-nonane) dimer, and particularly preferably9-BBN(9-borabicyclo[3.3.1]-nonane). The amount of the organic boranereagent used is not particularly limited insofar as an alkyl boraneintermediate can be produced. The organic borane reagent can be used inan amount of 1 to 20 moles per mole of the compound represented byFormula (XXII); the amount of the organic borane reagent is preferably 6to 10 moles from the viewpoint of facilitating the progress of thereaction.

As a transition metal catalyst, for example, a bivalent palladiumcatalyst (e.g., palladium acetate, palladium chloride, and1,1′-bis(diphenylphosphino)ferrocene-palladium (II) dichloride) can beused. If necessary, a ligand (e.g., triphenylphosphine andtri-tert-butylphosphine) can be used. The amount of the transition metalcatalyst used may vary depending on the type of catalyst. The transitionmetal catalyst is typically used in an amount of 0.0001 to 1 mole, andpreferably 0.01 to 0.5 moles, per mole of the compound represented byFormula (XXII). The ligand is typically used in an amount of 0.0001 to 4moles, and preferably 0.01 to 2 moles, per mole of the compoundrepresented by Formula (XXII).

Alternatively, for example, a zero-valent palladium catalyst can beused. Examples of zero-valent palladium catalysts includetetrakis(triphenylphosphine)palladium,tris(dibenzylideneacetone)dipalladium, palladium carbon, and the like.Tetrakis(triphenylphosphine)palladium ortris(dibenzylideneacetone)dipalladium is preferable, andtetrakis(triphenylphosphine)palladium is particularly preferable. Theamount of the zero-valent palladium catalyst used is not particularlylimited insofar as the intramolecular cyclization reaction can proceed,and may vary depending on the type of catalyst. The zero-valentpalladium catalyst can be used in an amount of 0.0001 to 1 mole, andpreferably 0.01 to 0.5 moles, per mole of the compound represented byFormula (XXII).

If necessary, a ligand may be added with a zero-valent palladiumcatalyst. Examples of such ligands include triphenylphosphine,1,1′-bis(diphenylphosphino)ferrocene, tri-tert-butylphosphine,tricyclohexylphosphine, 2-dicyclohexylphosphino-2′,6′-dimethoxybiphenyl,2-dicyclohexylphosphino-2′,4′,6′-triisopropylbiphenyl,2-(di-tert-butylphosphino)biphenyl,2-dicyclohexylphosphino-2′-(N,N-dimethylamino)biphenyl,4,5′-bis(diphenylphosphino)-9,9′-dimethylxanthene, and the like. Whentris(dibenzylideneacetone)dipalladium is used as a zero-valent palladiumcatalyst, triphenylphosphine can be added as a ligand. The amount of theligand used is not particularly limited insofar as the intramolecularcyclization reaction can proceed. The ligand can be used in an amount of0.0001 to 4 moles, and preferably 0.01 to 2 moles, per mole of thecompound represented by Formula (XXII).

Examples of bases include inorganic bases such as sodium hydrogencarbonate, sodium carbonate, potassium carbonate, cesium carbonate, andalkali metal hydroxides. Alkali metal hydroxides are preferable.Examples of alkali metal hydroxides include lithium hydroxide, sodiumhydroxide, potassium hydroxide, and cesium hydroxide. Lithium hydroxide,sodium hydroxide, potassium hydroxide, or cesium hydroxide is preferablyused. Lithium hydroxide or sodium hydroxide is particularly preferable.The amount of the base used is not particularly limited insofar as thereaction proceeds. The base can be used in an amount of 1 to 100 moles,and preferably 2 to 20 moles, per mole of the compound represented byFormula (XXII). Alkali metal hydroxide can be used in the form of anaqueous alkali metal hydroxide solution.

As the combination of an organic borane reagent, an alkali metalhydroxide, and a zero-valent palladium catalyst, a combination of apreferable organic borane reagent, a preferable alkali metal hydroxide,and a preferable zero-valent palladium catalyst is preferable. Acombination of a particularly preferable organic borane reagent, aparticularly preferable alkali metal hydroxide, and a particularlypreferable zero-valent palladium catalyst is particularly preferable.

Any solvent that does not adversely affect the reaction can be used.Examples thereof include hydrocarbons (e.g., benzene, toluene, andxylene), ethers (e.g., 1,2-dimethoxyethane, tetrahydrofuran, and1,4-dioxane), aprotic polar solvents (e.g., N,N-dimethylformamide,dimethyl sulfoxide, and hexamethyl phosphoryl amide), water, andmixtures thereof. 1,2-Dimethoxyethane or tetrahydrofuran is preferablyused. Tetrahydrofuran is particularly preferable from the viewpoint ofstability of the organic borane reagent and the generated alkylboraneintermediate. The amount of the solvent used is not particularly limitedinsofar as the reaction proceeds. The solvent can be used in an amountthat is 1 to 300 times, and preferably 10 to 96 times, the weight of thecompound represented by Formula (XXII).

The reaction time is not particularly limited insofar as the compound ofFormula (XXIII) can be obtained. The reaction time may be 0.1 to 100hours, and preferably 0.5 to 24 hours.

The reaction temperature is not particularly limited insofar as thecompound represented by Formula (XXIII) can ultimately be obtained. Thereaction temperature may be −20° C. to the boiling temperature of thesolvent, and preferably 0° C. to 150° C. In the intramolecularcyclization reaction of the alkylborane intermediate using a zero-valentpalladium catalyst and an alkali metal hydroxide aqueous solution, a lowreaction temperature tends to cause side reactions, which results in alow yield. Therefore, the temperature is preferably 61° C. or higher.

The thus-obtained compound represented by Formula (XXIII) can besubjected to the subsequent step after or without isolation orpurification by known isolation and purification means, such asconcentration, vacuum concentration, crystallization, solventextraction, reprecipitation, and chromatography.

In this step, generation of an alkylborane intermediate in the systemcan be confirmed. For example, LCMS spectra can be used as theconfirmation method.

-   Step w

In this step, the compound represented by Formula (XXIV) is produced byreacting the compound represented by Formula (XXIII) with iodobenzenediacetate.

The amount of the iodobenzene diacetate used in this step is 1 to 10moles, preferably 1 to 2 moles, per mole of the compound represented byFormula (XXIII).

In the above reaction, if necessary, tetrabutylammonium iodide may beadded. The tetrabutylammonium iodide can be added in an amount of 0.01to 10 moles, preferably 0.1 to 1 mole, per mole of the compoundrepresented by Formula (XXIII).

Any reaction solvent that does not adversely affect the reaction can beused. Examples of suitable reaction solvents include methylene chloride,chloroform, 1,2-dichloroethane, acetic acid and mixed solvents thereof.

The reaction temperature is generally 0° C. to the boiling temperatureof the solvent, and is preferably from 0° C. to room temperature. Thereaction time is 0.1 to 100 hours, preferably 0.1 to 24 hours.

The thus-obtained compound represented by Formula (XXIV) can besubjected to the subsequent step after or without isolation orpurification by known isolation and purification means, such asconcentration, vacuum concentration, crystallization, solventextraction, reprecipitation, and chromatography.

-   Step x

In this step, the protected hydroxy group of the compound represented byFormula (XXIV) is deprotected to produce the compound of Formula (XXV).

The deprotection can be performed by a known method, such as the methoddescribed in Protective Groups in Organic Synthesis, T. W. Greene, JohnWiley & Sons (1981); or a similar method.

When the acetyl group is deprotected, examples of the deprotectionreagent include sodium hydroxide, potassium hydroxide, and the like. Theamount of the reagent used is preferably 1 to 100 moles per mole of thecompound represented by Formula (XXIV).

Any solvent that does not adversely affect the reaction can be used.Examples of the solvent include water, methanol, ethanol,tetrahydrofuran, and mixed solvents thereof. The reaction time is 0.1 to100 hours, and preferably 0.5 to 24 hours. The reaction temperature is0° C. to the boiling temperature of the solvent, and is preferably from0 to 50° C.

The thus-obtained compound represented by Formula (XXV) can be subjectedto the subsequent step after or without isolation or purification byknown isolation and purification means, such as concentration, vacuumconcentration, crystallization, solvent extraction, reprecipitation, andchromatography.

-   Step y

In this step, the compound represented by Formula (XXVI) is produced bysubjecting the compound represented by Formula (XXV) to an eliminationreaction.

In the elimination reaction, acids such as p-toluenesulfonic acidmonohydrate, 10-camphorsulfonic acid, and the like are used. The amountof the acid used is 0.1 to 100 moles, preferably 1 to 10 moles, per moleof the compound represented by Formula (XXV).

Any solvent that does not adversely affect the reaction can be used.Examples of the solvent include toluene, xylene, and mixed solventsthereof. The reaction time is 0.1 to 100 hours, and preferably 0.5 to 24hours. The reaction temperature ranges from room temperature to theboiling temperature of the solvent.

The thus-obtained compound represented by Formula (XXVI) can besubjected to the subsequent step after or without isolation orpurification by known isolation and purification means, such asconcentration, vacuum concentration, crystallization, solventextraction, reprecipitation, and chromatography.

-   Step z

This step can be performed in the same manner as in Step g.

In the above production methods 1 to 6, for functional groups having anactive proton, such as amino, imino, hydroxy, carboxy, carbonyl, andamide groups, and indole, protected reagents can be used, or aprotecting group can be introduced into such a functional groupaccording to a usual method; afterward, the protecting group can beremoved in an appropriate step in each production method.

The “protecting group of an amino group or protecting group of an iminogroup” is not particularly limited, insofar as it has a protectingfunction. Examples of such protecting groups include aralkyl groups suchas benzyl, p-methoxybenzyl, 3,4-dimethoxybenzyl, o-nitrobenzyl,p-nitrobenzyl, benzhydryl, trityl, and cumyl; lower alkanoyl groups suchas formyl, acetyl, propionyl, butyryl, pivaloyl, trifluoroacetyl, andtrichloroacetyl; benzoyl; arylalkanoyl groups such as phenylacetyl andphenoxyacetyl; lower alkoxycarbonyl groups such as methoxycarbonyl,ethoxycarbonyl, propyloxycarbonyl, and tert-butoxycarbonyl;aralkyloxycarbonyl groups such as p-nitrobenzyloxycarbonyl andphenethyloxycarbonyl; lower alkylsilyl groups such as trimethylsilyl andtert-butyldimethylsilyl; tetrahydropyranyl; trimethylsilylethoxymethyl;lower alkylsulfonyl groups such as methylsulfonyl, ethylsulfonyl, andtert-butylsulfonyl; lower alkylsulfinyl groups such astert-butylsulfinyl; arylsulfonyl groups such as benzenesulfonyl andtoluenesulfonyl; and imido groups such as phthalimido. In particular,trifluoroacetyl, acetyl, tert-butoxycarbonyl, benzyloxycarbonyl,trimethylsilylethoxymethyl, cumyl, and the like are preferable.

The “protecting group of a hydroxy group” is not particularly limitedinsofar as it has a protecting function. Examples of such protectinggroups include lower alkyl groups such as methyl, ethyl, propyl,isopropyl, and tert-butyl; lower alkylsilyl groups such astrimethylsilyl and tert-butyldimethylsilyl; lower alkoxymethyl groupssuch as methoxymethyl and 2-methoxyethoxymethyl; tetrahydropyranyl;trimethylsilylethoxymethyl; aralkyl groups such as benzyl,p-methoxybenzyl, 2,3-dimethoxybenzyl, o-nitrobenzyl, p-nitrobenzyl, andtrityl; and acyl groups such as formyl, acetyl, and trifluoroacetyl. Inparticular, methyl, methoxymethyl, tetrahydropyranyl,trimethylsilylethoxymethyl, tert-butyldimethylsilyl, and acetyl arepreferable.

The “protecting group of a carboxy group” is not particularly limitedinsofar as it has a protecting function. Examples of such protectinggroups include lower alkyl groups such as methyl, ethyl, propyl,isopropyl, and tert-butyl; halo-lower-alkyl groups such as2,2,2-trichloroethyl; lower alkenyl groups such as allyl;trimethylsilylethoxymethyl; and aralkyl groups such as benzyl,p-methoxybenzyl, p-nitrobenzyl, benzhydryl, and trityl. In particular,methyl, ethyl, tert-butyl, allyl, benzyl, p-methoxybenzyl,trimethylsilylethoxymethyl, and the like are preferable.

The “protecting group of a carbonyl group” is not particularly limitedinsofar as it has a protecting function. Examples of such protectinggroups include ethylene ketal, trimethylene ketal, dimethyl ketal, andlike ketals and acetals.

The method for removing such a protecting group may vary depending onthe type of protecting group, stability of the desired compound (I),etc. For example, the following methods can be used: solvolysis using anacid or a base according to the method disclosed in a publication(Protective Groups in Organic Synthesis, third edition, T. W. Green,John Wiley & Sons (1999)) or a similar method, i.e., a method comprisingreacting with 0.01 moles or a large excess of an acid, preferablytrifluoroacetic acid, formic acid, or hydrochloric acid, or an equimolarto large excessive molar amount of a base, preferably potassiumhydroxide or calcium hydroxide; chemical reduction using a metal hydridecomplex, etc.; or catalytic reduction using a palladium-carbon catalyst,Raney nickel catalyst, etc.

The compound of the present invention can be isolated and purified byusual isolation and purification means. Examples of such means includesolvent extraction, recrystallization, preparative reversed-phasehigh-performance liquid chromatography, column chromatography,preparative thin-layer chromatography, and the like.

When the compound of the present invention has isomers such as opticalisomers, stereoisomers, regioisomers, and rotational isomers, any of theisomers and mixtures thereof is included within the scope of thecompound of the present invention. For example, when the compound hasoptical isomers, the optical isomer separated from a racemic mixture isalso included within the scope of the compound of the present invention.Each of such isomers can be obtained as a single compound by knownsynthesis and separation means (e.g., concentration, solvent extraction,column chromatography, recrystallization, etc.).

In the present invention, the carbon atom bound to a substituent B inFormula (I) is an asymmetric carbon; therefore, the compound includesisomers. As stated above, unless otherwise specified, the compound ofthe present invention includes all of the enantiomers and mixturesthereof. The compound of the present invention may be a mixture of R andS enantiomers. Such a mixture may be a mixture comprising 90% or more,95% or more, or 99% or more of R enantiomer; or a mixture comprising 90%or more, 95% or more, or 99% or more of S enantiomer.

Methods for chiral resolution include, for example: diastereomer methodof causing a chiral resolving agent to act on the compound of thepresent invention to form a salt, and resolving one of the enantiomersusing a solubility difference etc., of the obtained salt; preferentialcrystallization method of adding one of the enantiomers to asupersaturated solution of a racemate as a seed for crystallization; andcolumn chromatography such as HPLC using a chiral column. A chiralresolving agent that can be used in the diastereomer method can beappropriately selected from, for example, acid resolving agents such astartaric acid, malic acid, lactic acid, mandelic acid,10-camphorsulfonic acid, and derivatives thereof; and basic resolvingagents such as brucine, strychnine, quinine, and like alkaloidcompounds, amino acid derivatives, cinchonidine, andα-methylbenzylamine. In addition, one of the enantiomers of the compoundof the present invention alone can be obtained not only by obtaining thecompound of the present invention as a mixture of each of theenantiomers and then conducting the above described methods of chiralresolution, but also by obtaining, through chiral resolution by theabove described methods etc., and using one enantiomer of the compoundof the present invention as a synthetic raw material. Furthermore,methods for obtaining one of the enantiomers of the compound of thepresent invention or its raw material compound include a method ofpreferentially obtaining one of the enantiomers by adjusting reactionconditions for a catalyst or the like in a reaction step of generatingasymmetric carbon.

The compound of the present invention or a salt thereof may be in theform of crystals. Single crystals and polymorphic mixtures are includedwithin the scope of the compound of the present invention or a saltthereof. Such crystals can be produced by crystallization according to acrystallization method known per se in the art. The compound of thepresent invention or a salt thereof may be a solvate (e.g., a hydrate)or a non-solvate. Any of such forms are included within the scope of thecompound of the present invention or a salt thereof. Compounds labeledwith an isotope (e.g., 3H, 14C, 35S, and 125I) are also included withinthe scope of the compound of the present invention or a salt thereof.

The salt of the compound of the present invention or of the intermediatethereof refers to a common salt used in the field of organic chemistry.Examples of such salts include base addition salts to carboxy when thecompound has carboxy, and acid addition salts to an amino or basicheterocyclic group when the compound has an amino or basic heterocyclicgroup.

Examples of base addition salts include alkali metal salts such assodium salts and potassium salts; alkaline earth metal salts such ascalcium salts and magnesium salts; ammonium salts; and organic aminesalts such as trimethylamine salts, triethylamine salts,dicyclohexylamine salts, ethanolamine salts, diethanolamine salts,triethanolamine salts, procaine salts, and N,N′-dibenzylethylenediaminesalts.

Examples of acid addition salts include inorganic acid salts such ashydrochlorides, sulfates, nitrates, phosphates, and perchlorates;organic acid salts such as acetates, formates, maleates, fumarates,tartrates, citrates, ascorbates, and trifluoroacetates; and sulfonatessuch as methanesulfonates, isethionates, benzenesulfonates, andp-toluenesulfonates.

The compound of the present invention or a salt thereof has excellentEGFR inhibitory activity and is useful as an antitumor agent. Further,the compound of the present invention or a salt thereof has excellentselectivity toward EGFR, and advantageously fewer side effects caused byother kinases. The type of malignant tumor to be treated by the compoundof the present invention or a salt thereof is not particularly limited.Examples of malignant tumors include epithelial cancers (e.g.,respiratory system cancers, digestive system cancers, reproductivesystem cancers, secretion system cancers, and the like), sarcomas,hematopoietic tumors, central nervous system tumors, and peripheralnerve tumors. Preferable examples include epithelial cancers. Morepreferable examples include respiratory system cancers. Further, theorgan from which the tumor is developed is not particularly limited.Examples include head and neck cancers, esophagus cancer, gastriccancer, colon cancer, rectum cancer, liver cancer, gallbladder cancer,cholangiocarcinoma, biliary tract cancer, pancreatic cancer, lungcancer, breast cancer, ovarian cancer, cervical cancer, endometrialcancer, renal cancer, bladder cancer, prostate cancer, testicular tumor,osteosarcoma, soft-tissue sarcoma, blood cancer, multiple myeloma, skincancer, brain tumor, and mesothelioma. Preferably, the target cancer ishead and neck cancers, gastric cancer, colon cancer, rectum cancer,liver cancer, pancreatic cancer, lung cancer, breast cancer, ovariancancer, renal cancer, or prostate cancer, particularly preferably lungcancer.

Further, the compound of the present invention or a salt thereof hasexcellent inhibitory activity against mutated EGFR. Examples of such amutated EGFR include drug-tolerant mutated EGFR and hypersensitivemutated EGFR. Therefore, the compound of the present invention or a saltthereof is useful as an antitumor agent for treating the above malignanttumors having mutated EGFR.

When the compound of the present invention or a salt thereof is used asa pharmaceutical preparation, a pharmaceutical carrier can be added, ifrequired, thereby forming a suitable dosage form according to preventionand treatment purposes. Examples of the dosage form include oralpreparations, injections, suppositories, ointments, patches, and thelike. Of these, oral preparations are preferable. Such dosage forms canbe formed by methods conventionally known to persons skilled in the art.

As the pharmaceutical carrier, various conventional organic or inorganiccarrier materials used as preparation materials may be blended as anexcipient, binder, disintegrant, lubricant, or colorant in solidpreparations; or as a solvent, solubilizing agent, suspending agent,isotonizing agent, buffer, or soothing agent in liquid preparations.Moreover, pharmaceutical preparation additives, such as antiseptics,antioxidants, colorants, sweeteners, and stabilizers, may also be used,if required.

Oral solid preparations are prepared as follows. After an excipient isadded optionally with a binder, disintegrant, lubricant, colorant,taste-masking or flavoring agent, etc., to the compound of the presentinvention, the resulting mixture is formulated into tablets, coatedtablets, granules, powders, capsules, or the like by ordinary methods.

Examples of excipients include lactose, sucrose, D-mannitol, glucose,starch, calcium carbonate, kaolin, microcrystalline cellulose, andsilicic acid anhydride. Examples of binders include water, ethanol,1-propanol, 2-propanol, simple syrup, liquid glucose, liquid α-starch,liquid gelatin, D-mannitol, carboxymethyl cellulose, hydroxypropylcellulose, hydroxypropyl starch, methyl cellulose, ethyl cellulose,shellac, calcium phosphate, polyvinylpyrrolidone, and the like. Examplesof disintegrators include dry starch, sodium alginate, powdered agar,sodium hydrogen carbonate, calcium carbonate, sodium lauryl sulfate,stearic acid monoglyceride, lactose, and the like. Examples oflubricants include purified talc, stearic acid salt sodium, magnesiumstearate, borax, polyethylene glycol, and the like. Examples ofcolorants include titanium oxide, iron oxide, and the like. Examples oftaste-masking or flavoring agents include sucrose, bitter orange peel,citric acid, tartaric acid, and the like.

When a liquid preparation for oral administration is prepared, ataste-masking agent, a buffer, a stabilizer, a flavoring agent, and thelike may be added to the compound of the present invention; and theresulting mixture may be formulated into an oral liquid preparation,syrup, elixir, etc., according to an ordinary method.

In this case, the same taste-masking or flavoring agent as thosementioned above may be used. An example of the buffer is sodium citrate,and examples of the stabilizer include tragacanth, gum arabic, andgelatin. As necessary, these preparations for oral administration may becoated according to methods known in the art with an enteric coating orother coating for the purpose of, for example, persistence of effects.Examples of such coating agents include hydroxypropyl methylcellulose,ethyl cellulose, hydroxymethyl cellulose, hydroxypropyl cellulose,polyoxyethylene glycol, and Tween 80(registered trademark).

When an injection agent is prepared, a pH regulator, a buffer, astabilizer, an isotonizing agent, a local anesthetic, and the like, maybe added to the compound of the present invention; and the mixture maybe formulated into a subcutaneous, intramuscular, or intravenousinjection according to an ordinary method.

Examples of the pH adjuster and the buffer used herein include sodiumcitrate, sodium acetate, and sodium phosphate. Examples of thestabilizer include sodium pyrosulfite, EDTA, thioglycolic acid, andthiolactic acid. Examples of the local anesthetic include procainehydrochloride and lidocaine hydrochloride. Examples of the tonicityagent include sodium chloride, dextrose, D-mannitol, and glycerol.

When a suppository is prepared, pharmaceutically acceptable carriersknown in the art, such as polyethylene glycol, lanolin, cacao butter,and fatty acid triglyceride; and as necessary, surfactants such as Tween80 (registered trademark), may be added to the compound of the presentinvention, and the resulting mixture may be formulated into asuppository according to an ordinary method.

When an ointment is prepared, a commonly used base, stabilizer, wettingagent, preservative, and the like, may be blended into the compound ofthe present invention, as necessary; and the obtained mixture may bemixed and formulated into an ointment according to an ordinary method.

Examples of the base include liquid paraffin, white petrolatum, whitebeeswax, octyl dodecyl alcohol, and paraffin.

Examples of the preservative include methyl paraoxybenzoate, ethylparaoxybenzoate, and propyl paraoxybenzoate.

When a patch is prepared, the above-described ointment, cream, gel,paste, or the like, may be applied to an ordinary substrate according toan ordinary method.

As the substrate, woven fabrics or non-woven fabrics comprising cotton,staple fibers, or chemical fibers; and films or foam sheets of softvinyl chloride, polyethylene, polyurethane, etc., are suitable.

The amount of the compound of the present invention to be incorporatedin each of such dosage unit forms depends on the condition of thepatient to whom the compound is administered, the dosage form thereof,etc. In general, in the case of an oral agent, the amount of thecompound is 0.05 to 1000 mg per dosage unit form. In the case of aninjection, the amount of the compound is 0.01 to 500 mg per dosage unitform; and in the case of a suppository, the amount of the compound is 1to 1000 mg per dosage unit form.

The daily dose of the medicine in such a dosage form depends on thecondition, body weight, age, gender, etc., of the patient, and cannot begeneralized. For example, the daily dose for an adult (body weight: 50kg) may be generally 0.05 to 5,000 mg, and preferably 0.1 to 1,000 mg;and is preferably administered in one dose, or in two to three divideddoses, per day.

Examples of mammals to which the compound of the present invention isadministered include humans, monkeys, mice, rats, rabbits, dogs, cats,cows, horses, pigs, and sheep.

EXAMPLES

The present invention is more specifically explained below withreference to Examples; however, the present invention is not limited tothese Examples.

In the Examples, commercially available reagents were used, unlessotherwise specified. Purif-Pack® SI, produced by Moritex Corp. (producedby Shoko Scientific Co., Ltd.); KP-Sil® Silica prepacked column,produced by Biotage; HP-Sil(r) Silica prepacked column, produced byBiotage, or HP-Sphere® Silica prepacked column, produced by Biotage, wasused for the silica gel column chromatography. Purif-Pack® NH, producedby Moritex Corp. (produced by Shoko Scientific Co., Ltd.); or KP-NH®prepacked column, produced by Biotage, was used for the basic silica gelcolumn chromatography. Kieselgel TM 60F254, Art. 5744, produced byMerck, or NH2 Silica Gel 60F254 Plate, produced by Wako, was used forthe preparative thin-layer chromatography. NMR spectrum was measured byusing AL400 (400 MHz; produced by JEOL), or Mercury 400 (400 MHz;produced by Agilent Technologies, Inc.) spectrometer. When thedeuterated solvent contains tetramethylsilane, tetramethylsilane wasused as the internal reference. Otherwise, an NMR solvent was used asthe internal reference. All of the δ values are shown by ppm. Themicrowave reaction was performed using an Initiator, produced byBiotage.

Further, the LCMS spectrum was measured using an Acquity SQD(quadrupole), produced by Waters Corporation, under the followingconditions.

-   Column: Acquity UPLC® BEH C18, 2.1×50 mm, 1.7 μm, produced by Waters    Corporation-   MS detection: ESI positive-   UV detection: 254 and 210 nm-   Column flow rate: 0.5 mL/min-   Mobile phase: Water/acetonitrile (0.1% formic acid)-   Injection volume: 1 μL-   Gradient (Table 1)

Time (min) Water Acetonitrile 0 95 5 0.1 95 5 2.1 5 95 3.0 STOP.

Reversed-phase preparative HPLC purification was performed using apreparative separation system available from Gilson.

-   Column: CombiPrep Pro C18, 50×30 mm I.D., S-5 μm (produced by YMC).-   UV detection: 254 nm-   Column flow rate: 40 mL/min-   Mobile phase: Water/acetonitrile (0.1% trifluoroacetic acid)-   Injection volume: 0.1 to 1 mL.

Each symbol stands for the following.

-   s: Singlet-   d: Doublet-   t: Triplet-   q: Quartet-   dd: Double Doublet-   dt: Double Triplet-   ddd: Double Doublet-   m: Multiplet-   brs: Broad Singlet-   DMSO-d₆: Deuterated dimethyl sulfoxide-   CDCl₃: Deuterated chloroform-   CD₃OD: Deuterated methanol-   THF: Tetrahydrofuran-   DMF: N,N-dimethylformamide-   DME: 1,2-Dimethoxyethane-   DMSO: Dimethylsulfoxide-   HATU: O-(7-azabenzotriazol-1-yl)-N,N,N′,N′-tetramethyluronium    hexafluorophosphate

Example 1(S)—N-(4-amino-6-methylene-5-(quinolin-3-yl)-7,8-dihydro-6H-pyrimido[5,4-b]pyrrolizin-7-yl)acrylamide(Compound 1)

Step 1: Synthesis of (S)-tert-butyl(1-(4-chloro-5-iodo-7H-pyrrolo[2,3-d]pyrimidin-7-yl)but-3-en-2-yl)carbamate

Diisopropyl azodicarboxylate (2.44 ml) was slowly added to a solution oftriphenylphosphine (13.1 g) in THF (70 ml) under ice-cooling. Thereaction mixture was stirred under ice-cooling for 1 hour, and then asolution of (S)-tert-butyl (1-hydroxybut-3-en-2-yl)carbamate (7.0 g)synthesized according to the method disclosed in NPD Org. Lett., 2005,vol. 7, No. 5, pp. 847-849 and4-chloro-5-iodo-7H-pyrrolo[2,3-d]pyrimidine (6.97 g) in THF (35 ml) wasslowly added thereto. After the reaction mixture was stirred at roomtemperature for 2 hours, the solvent was distilled off under reducedpressure. The resulting residue was purified by silica gel columnchromatography (developing solvent: hexane/ethyl acetate), therebyobtaining the title compound (20.84 g) as a light-yellow, oilysubstance.

ESI-MS m/z 448, 450 [M+H]⁺.

Step 2: Synthesis of (S)-tert-butyl(1-(4-amino-5-iodo-7H-pyrrolo[2,3-d]pyrimidin-7-yl)but-3-en-2-yl)carbamate

An 8 N ammonia-methanol solution (89.4 ml) was added to the(S)-tert-butyl(1-(4-chloro-5-iodo-7H-pyrrolo[2,3-d]pyrimidin-7-yl)but-3-en-2-yl)carbamate(20.84 g) obtained in Step 1, and the mixture was stirred in anautoclave at 120° C. for 6 hours. The reaction mixture was cooled withice, and the solvent was distilled off under reduced pressure. After theresulting residue was diluted with a small amount of methanol, theresulting precipitate was collected by filtration, washed with coldmethanol (11 ml), and then dried under reduced pressure, therebyobtaining the title compound (8.28 g) as a milky-white solid.

ESI-MS m/z 430 [M+H]⁺.

Step 3: Synthesis of (S)-tert-butyl(1-(4-amino-5-(quinolin-3-yl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)but-3-en-2-yl)carbamate

A mixture of the (S)-tert-butyl(1-(4-amino-5-iodo-7H-pyrrolo[2,3-d]pyrimidin-7-yl)but-3-en-2-yl)carbamate(8.26 g) obtained in Step 2, 3-quinolineboronic acid (4.99 g), cesiumcarbonate (12.54 g),1,1′-bis(diphenylphosphino)ferrocene-palladium(II)dichloride (785.6 mg),DME (66 ml), and water (33 ml) was stirred under a nitrogen atmosphereat 100° C. for 2 hours. After cooling the reaction mixture, water andethyl acetate were added thereto to separate the organic layer. Theaqueous layer was then extracted with ethyl acetate twice. The resultingorganic layer was dried over anhydrous magnesium sulfate, and thesolvent was distilled off under reduced pressure. The resulting residuewas purified by silica gel column chromatography (developing solvent:hexane/ethyl acetate, ethyl acetate/methanol), thereby obtaining thetitle compound (8.0 g) as a light-orange solid.

ESI-MS m/z 431 [M+H]⁺.

Step 4: Synthesis of (S)-tert-butyl(1-(4-amino-6-bromo-5-(quinolin-3-yl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)but-3-en-2-yl)carbamate

N-Bromosuccinimide (3.63 g) was added to a solution of the(S)-tert-butyl(1-(4-amino-5-(quinolin-3-yl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)but-3-en-2-yl)carbamate(7.98 g) obtained in Step 3 in DMF (64 ml) at −15° C., and the mixturewas stirred at −15° C. for 1 hour. A 10% aqueous sodium thiosulfatesolution and ethyl acetate were added to the reaction mixture, andstirred at room temperature for 10 minutes. The organic layer wasseparated, and the aqueous layer was extracted with ethyl acetate twice.The resulting organic layer was washed with a saturated sodium chloridesolution twice, and dried over anhydrous magnesium sulfate. The solventwas distilled off under reduced pressure. The resulting residue waspurified by silica gel column chromatography (developing solvent: ethylacetate/methanol), thereby obtaining the title compound (6.30 g) as alight-brown solid.

ESI-MS m/z 509, 511 [M+H]⁺.

Step 5: Synthesis of (S)-tert-butyl(4-amino-6-methylene-5-(quinolin-3-yl)-7,8-dihydro-6H-pyrimido[5,4-b]pyrrolizin-7-yl)carbamate

4N sodium hydroxide aqueous solution (28.8 ml) was added to a solutionof the (S)-tert-butyl(1-(4-amino-6-bromo-5-(quinolin-3-yl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)but-3-en-2-yl)carbamate(5.00 g) obtained in Step 4 in THF (100 ml), and the mixture wasdegassed under reduced pressure, followed by nitrogen purging. Aftertetrakis(triphenylphosphine)palladium (1.13 g) was added, the mixturewas stirred overnight while being heated under reflux. After thereaction mixture was cooled to room temperature, extraction wasperformed using ethyl acetate, and the resulting organic layer waswashed with a saturated sodium chloride solution and dried overanhydrous sodium sulfate. After filtration and concentration, theresulting residue was purified by silica gel chromatography (developingsolvent: ethyl acetate/methanol), thereby obtaining the title compoundas an orange solid (5.01 g).

ESI-MS m/z 429 [M+H]⁺.

Step 6: Synthesis of Compound 1

A 4 N hydrogen chloride-dioxane solution (1 ml) and methanol (1 ml) wereadded to a solution of the (S)-tert-butyl(4-amino-6-methylene-5-(quinolin-3-yl)-7,8-dihydro-6H-pyrimido[5,4-b]pyrrolizin-7-yl)carbamate(100 mg) obtained in Step 5 in chloroform (1 ml), and the mixture wasstirred for 5 hours at room temperature. The reaction mixture wasconcentrated under reduced pressure, thereby yielding(S)-6-methylene-5-(quinolin-3-yl)-7,8-dihydro-6H-pyrimido[5,4-b]pyrrolizin-4,7-diaminehydrochloride.

Diisopropylethylamine (0.187 ml) was added to a solution of(S)-6-methylene-5-(quinolin-3-yl)-7,8-dihydro-6H-pyrimido[5,4-b]pyrrolizin-4,7-diaminehydrochloride in chloroform (4 ml), and a 100 mg/ml solution of acryloylchloride in chloroform (0.19 ml) was added under ice-cooling. Themixture was stirred for 40 minutes. The reaction mixture was purified bysilica gel chromatography (developing solvent: ethyl acetate/methanol);after concentration, the resulting residue was suspended and washed inethyl acetate/hexane. The resulting solid was collected by filtrationand dried under reduced pressure, thereby obtaining the title compound(49 mg) as a light-green solid.

¹H-NMR (DMSO-d₆)δ: 3.88-3.93 (1H, m), 4.57-4.63 (1H, m), 5.03 (1H, d,J=2.4 Hz), 5.24 (1H, d, J=2.4 Hz), 5.55-5.62 (1H, m), 5.68 (1H, dd,J=10.0, 2.4 Hz), 6.12-6.38 (4H, m), 7.65 (1H, dd, J=7.8, 7.8 Hz),7.77-7.83 (1H, m), 8.04-8.11 (2H, m), 8.15 (1H, s), 8.41 (1H, d, J=2.2Hz), 8.82 (1H, d, J=7.8 Hz), 8.98 (1H, d, J=2.2 Hz).

ESI-MS m/z 383[M+H]^(|).

Example 2 (S)—N-(4-amino-6-methylene-5-(quinolin-3-yl)-7,8-dihydro-6H-pyrimido[5,4-b]pyrrolizin-7-yl)methacrylamide (Compound 2)

Step 1: Synthesis of(S)-6-methylene-5-(quinolin-3-yl)-7,8-dihydro-6H-pyrimido[5,4-b]pyrrolizin-4,7-diamine

5 N hydrochloric acid (12 ml) was added to a solution of the(S)-tert-butyl(4-amino-6-methylene-5-(quinolin-3-yl)-7,8-dihydro-6H-pyrimido[5,4-b]pyrrolizin-7-yl)carbamate(5.01 g) obtained in Step 5 of Example 1 in ethanol (25 ml), and theresulting mixture was stirred for 1 hour at an external temperature of70 to 80° C. After the reaction mixture was cooled to room temperature,the reaction mixture was washed with chloroform, and the aqueous layerwas adjusted to about pH 10 using a 5 N sodium hydroxide aqueoussolution, followed by extraction with chloroform. The resulting organiclayer was dried over anhydrous sodium sulfate, followed by filtrationand concentration. The resulting residue was purified by basic silicagel chromatography (developing solvent: chloroform/methanol), therebyobtaining the title compound (2.10 g) as a yellow solid.

ESI-MS m/z 329 [M+H]⁺.

Step 2: Synthesis of Compound 2

Diisopropylethylamine (0.0318 ml) and methacryloyl chloride (0.0148 ml)were successively added to a solution of the(S)-6-methylene-5-(quinolin-3-yl)-7,8-dihydro-6H-pyrimido[5,4-b]pyrrolizin-4,7-diamine(50.0 mg) obtained in Step 1 in acetonitrile (2.0 ml) and water (2.0 ml)at 0° C., and the resulting mixture was stirred for 45 minutes at thesame temperature. The reaction mixture was poured into a saturatedaqueous sodium bicarbonate solution, followed by extraction withchloroform. After the organic layer was dried over anhydrous sodiumsulfate, the solvent was distilled off under reduced pressure. Theresulting residue was purified by silica gel column chromatography(developing solvent: ethyl acetate/methanol), thereby obtaining thetitle compound (38.0 mg) as a light-yellow solid.

¹H-NMR (CDCl₃)δ: 2.02 (3H, s), 4.03 (1H, dd, J=11.6, 4.9 Hz), 4.70 (1H,dd, J=11.6, 8.2 Hz), 5.20-5.22 (3H, m), 5.44 (2H, d, J=1.7 Hz),5.73-5.75 (1H, m), 5.82 (1H, s), 6.89 (1H, d, J=7.6 Hz), 7.61-7.65 (1H,m), 7.76-7.80 (1H, m), 7.87 (1H, d, J=8.3 Hz), 8.15 (1H, d, J=8.5 Hz),8.21-8.25 (2H, m), 9.06 (1H, d, J=2.2 Hz).

ESI-MS m/z 397[M+H]⁺.

Example 3 (S)—N-(4-amino-6-methylene-5-(quinolin-3-yl)-7,8-dihydro-6H-pyrimido[5,4-b]pyrrolizin-7-yl)but-2-enamide (mixture of E and Z)(Compound 3)

The same synthesis as in Step 2 of Example 2 was performed usingcrotonoyl chloride instead of the methacryloyl chloride used in Step 2of Example 2, thereby obtaining the title mixture (8.8 mg) as alight-yellow solid.

¹H-NMR (CDCl₃)δ: 1.83-1.90 (3H, m), 4.01 (1H, dd, J=11.6, 5.2 Hz), 4.73(1H, dd, J=11.6, 8.2 Hz), 5.00 (2H, s), 5.19 (1H, d, J=1.5 Hz), 5.44(1H, d, J=2.0 Hz), 5.75-5.77 (1H, m), 5.87-5.91 (1H, m), 6.27 (1H, d,J=8.3 Hz), 6.92-7.01 (1H, m), 7.62-7.66 (1H, m), 7.78-7.82 (1H, m), 7.88(1H, d, J=7.3 Hz), 8.18 (1H, d, J=8.0 Hz), 8.28-8.29 (2H, m), 9.08 (1H,d, J=2.2 Hz).

ESI-MS m/z 397[M+H]⁺.

Example 4(S,E)-N-(4-amino-6-methylene-5-(quinolin-3-yl)-7,8-dihydro-6H-pyrimido[5,4-b]pyrrolizin-7-yl)-4-(dimethylamino)but-2-enamide(Compound 4)

(E)-4-(dimethylamino)but-2-enoic acid hydrochloride (60.6 mg), HATU (139mg), diisopropylethylamine (0.106 ml), and DMF (1.0 ml) weresuccessively added to a suspension of the(S)-6-methylene-5-(quinolin-3-yl)-7,8-dihydro-6H-pyrimido[5,4-b]pyrrolidin-4,7-diamine(100 mg) obtained in Step 1 of Example 2 in methylene chloride (3.0 ml)at room temperature, and the mixture was stirred for 1.5 hours at thesame temperature. The reaction mixture was poured into a saturatedaqueous sodium bicarbonate solution, followed by extraction with ethylacetate. After the organic layer was dried over anhydrous sodiumsulfate, the solvent was distilled off under reduced pressure. Theresulting residue was purified by silica gel column chromatography(developing solvent: ethyl acetate/methanol), thereby obtaining thetitle compound (114 mg) as a light-yellow solid.

¹H-NMR (CDCl₃)δ: 2.23 (6H, s), 3.07 (2H, dd, J=6.0, 1.3 Hz), 4.01 (1H,dd, J=11.6, 5.0 Hz), 4.70 (1H, dd, J=11.6, 8.2 Hz), 5.09 (2H, brs), 5.20(1H, d, J=2.0 Hz), 5.43 (1H, d, J=2.0 Hz), 5.73-5.79 (1H, m), 6.07 (1H,dt, J=15.4, 1.3 Hz), 6.72 (1H, brs), 6.94 (1H, dt, J=15.4, 6.0 Hz),7.61-7.65 (1H, m), 7.77-7.81 (1H, m), 7.87 (1H, d, J=8.0 Hz), 8.16 (1H,d, J=8.5 Hz), 8.25-8.26 (2H, m), 9.05 (1H, d, J=2.0 Hz).

ESI-MS m/z 440[M+H]⁺.

Example 5(S,E)-N-(4-amino-6-methylene-5-(quinolin-3-yl)-7,8-dihydro-6H-pyrimido[5,4-b]pyrrolizin-7-yl)-3-chloroacrylamide(Compound 5)

The same synthesis as in Example 4 was performed usingtrans-3-chloroacrylic acid instead of the(E)-4-(dimethylamino)but-2-enoic acid hydrochloride used in Example 4,thereby obtaining the title compound (49.2 mg) as a yellow solid.

¹H-NMR (CDCl₃)δ: 4.04 (1H, dd, J=11.7, 4.6 Hz), 4.69 (1H, dd, J=11.7,8.0 Hz), 5.03 (2H, s), 5.22 (1H, d, J=1.7 Hz), 5.44 (1H, d, J=1.7 Hz),5.72-5.75 (1H, m), 6.34 (1H, d, J=13.0 Hz), 6.89 (1H, brs), 7.41 (1H, d,J=13.0 Hz), 7.63-7.65 (1H, m), 7.79-7.81 (1H, m), 7.87 (1H, d, J=8.0Hz), 8.16 (1H, d, J=8.3 Hz), 8.25-8.26 (2H, m), 9.02 (1H, d, J=2.0 Hz).

ESI-MS m/z 417, 419[M+H]⁻.

Example 6(S,Z)—N-(4-amino-6-methylene-5-(quinolin-3-yl)-7,8-dihydro-6H-pyrimido[5,4-b]pyrrolizin-7-yl)-3-chloroacrylamide(Compound 6)

The same synthesis as in Example 4 was performed usingcis-3-chloroacrylic acid instead of the (E)-4-(dimethylamino)but-2-enoicacid hydrochloride used in Example 4, thereby obtaining the titlecompound (74.7 mg) as a yellow solid.

¹H-NMR (CDCl₃)δ: 4.07 (1H, dd, J=11.6, 5.4 Hz), 4.82 (1H, dd, J=11.6,8.2 Hz), 4.96 (2H, s), 5.26 (1H, d, J=2.0 Hz), 5.48 (1H, d, J=2.0 Hz),5.75-5.78 (1H, m), 6.28 (1H, d, J=8.3 Hz), 6.61 (1H, d, J=8.3 Hz), 6.81(1H, d, J=7.3 Hz), 7.64-7.67 (1H, m), 7.79-7.83 (1H, m), 7.90 (1H, d,J=7.1 Hz), 8.19 (1H, d, J=8.5 Hz), 8.31 (1H, d, J=2.0 Hz), 8.33 (1H, s),9.12 (1H, d, J=2.2 Hz).

ESI-MS m/z 417, 419[M+H]⁻.

Example 7(S,E)-N-(4-amino-6-methylene-5-(quinolin-3-yl)-7,8-dihydro-6H-pyrimido[5,4-b]pyrrolizin-7-yl)-4-(piperidin-1-yl)but-2-enamide(Compound 7)

The same synthesis as in Example 4 was performed using(E)-4-(piperidin-1-yl)but-2-enoic acid hydrochloride instead of the(E)-4-(dimethylamino)but-2-enoic acid hydrochloride used in Example 4,thereby obtaining the title compound (122 mg) as a yellow solid.

¹H-NMR (CDCl₃)δ: 1.42-1.59 (5H, m), 2.14 (1H, s), 2.39 (4H, brs), 3.10(2H, dd, J=6.0, 1.4 Hz), 4.01 (1H, dd, J=11.6, 5.0 Hz), 4.69 (1H, dd,J=11.5, 8.0 Hz), 5.10 (2H, brs), 5.20 (1H, d, J=2.1 Hz), 5.43 (1H, d,J=2.1 Hz), 5.73-5.79 (1H, m), 6.06 (1H, dt, J=15.3, 1.4 Hz), 6.76 (1H,brs), 6.96 (1H, dt, J=15.3, 6.0 Hz), 7.62-7.64 (1H, m), 7.77-7.80 (1H,m), 7.87 (1H, d, J=8.0 Hz), 8.16 (1H, d, J=8.3 Hz), 8.24-8.25 (2H, m),9.04 (1H, d, J=2.0 Hz).

ESI-MS m/z 480[M+H]⁺.

Example 8(S)—N-(4-amino-6-methylene-5-(quinolin-3-yl)-7,8-dihydro-6H-pyrimido[5,4-b]pyrrolizin-7-yl)propiolamide(Compound 8)

The same synthesis as in Example 4 was performed using propiolic acidinstead of the (E)-4-(dimethylamino)but-2-enoic acid hydrochloride usedin Example 4, thereby obtaining the title compound (30.0 mg) as a yellowsolid.

¹H-NMR (CDCl₃)δ: 2.91 (1H, s), 4.08 (1H, dd, J=11.7, 4.9 Hz), 4.76 (1H,dd, J=11.7, 8.0 Hz), 4.91 (2H, s), 5.24 (1H, d, J=1.2 Hz), 5.49 (1H, d,J=1.7 Hz), 5.68-5.69 (1H, m), 6.34-6.37 (1H, m), 7.65-7.67 (1H, m),7.81-7.83 (1H, m), 7.90 (1H, d, J=8.3 Hz), 8.20 (1H, d, J=8.5 Hz), 8.30(1H, d, J=2.0 Hz), 8.34 (1H, s), 9.11 (1H, d, J=2.2 Hz).

ESI-MS m/z 381[M+H]⁺.

Example 9(S)—N-(4-amino-6-methylene-5-(quinolin-3-yl)-7,8-dihydro-6H-pyrimido[5,4-b]pyrrolizin-7-yl)but-2-ynamide(Compound 9)

The same synthesis as in Example 4 was performed using but-2-ynoic acidinstead of the (E)-4-(dimethylamino)but-2-enoic acid hydrochloride usedin Example 4, thereby obtaining the title compound (98.0 mg) as a yellowsolid.

¹H-NMR (CDCl₃)δ: 1.96 (3H, s), 4.02 (1H, dd, J=11.6, 5.0 Hz), 4.70 (1H,dd, J=11.6, 8.3 Hz), 5.05 (2H, s), 5.24 (1H, d, J=1.6 Hz), 5.45 (1H, d,J=1.6 Hz), 5.70-5.72 (1H, m), 6.95-7.01 (1H, brs), 7.63-7.67 (1H, m),7.78-7.83 (1H, m), 7.88 (1H, d, J=8.3 Hz), 8.21 (1H, d, J=8.5 Hz),8.26-8.28 (2H, m), 9.02 (1H, d, J=2.0 Hz).

ESI-MS m/z 395[M+H]⁺.

Example 10(S,E)-N-(4-amino-6-methylene-5-(quinolin-3-yl)-7,8-dihydro-6H-pyrimido[5,4-b]pyrrolizin-7-yl)-4-(diethylamino)but-2-enamide(Compound 10)

The same synthesis as in Example 4 was performed using(E)-4-(diethylamino)but-2-enoic acid hydrochloride instead of the(E)-4-(dimethylamino)but-2-enoic acid hydrochloride used in Example 4,thereby obtaining the title compound (38.2 mg) as a yellow solid.

¹H-NMR (CDCl₃)δ: 1.00 (6H, t, J=7.2 Hz), 2.51 (4H, q, J=7.2 Hz), 3.22(2H, dd, J=5.9, 1.5 Hz), 4.00 (1H, dd, J=11.5, 4.9 Hz), 4.62 (1H, dd,J=11.5, 8.2 Hz), 5.22-5.24 (3H, m), 5.40 (1H, d, J=2.0 Hz), 5.74-5.80(1H, m), 6.12 (1H, dt, J=15.4, 1.5 Hz), 6.98 (1H, dt, J=15.4, 5.9 Hz),7.50 (1H, brs), 7.58-7.62 (1H, m), 7.74-7.78 (1H, m), 7.84 (1H, d, J=7.6Hz), 8.12-8.14 (2H, m), 8.22 (1H, d, J=1.7 Hz), 8.99 (1H, d, J=2.0 Hz).

ESI-MS m/z 468[M+H]⁺.

Example 11(S,E)-N-(4-amino-6-methylene-5-(quinolin-3-yl)-7,8-dihydro-6H-pyrimido[5,4-b]pyrrolizin-7-yl)-4-(ethyl(methyl)amino)but-2-enamide(Compound 11)

The same synthesis as in Example 4 was performed using(E)-4-(ethyl(methyl)amino)but-2-enoic acid hydrochloride instead of the(E)-4-(dimethylamino)but-2-enoic acid hydrochloride used in Example 4,thereby obtaining the title compound (13.5 mg) as a yellow solid.

¹H-NMR (CDCl₃)δ: 1.06 (3H, t, J=7.2 Hz), 2.24 (3H, s), 2.44 (2H, q,J=7.2 Hz), 3.15 (2H, dd, J=6.0, 1.6 Hz), 4.03 (1H, dd, J=11.6, 5.1 Hz),4.73 (1H, dd, J=11.6, 8.2 Hz), 5.01 (2H, s), 5.20 (1H, d, J=1.7 Hz),5.44 (1H, d, J=1.7 Hz), 5.75-5.77 (1H, m), 6.05 (1H, dt, J=15.4, 1.6Hz), 6.42 (1H, d, J=6.8 Hz), 6.96 (1H, dt, J=15.4, 6.0 Hz), 7.62-7.66(1H, m), 7.78-7.82 (1H, m), 7.88 (1H, d, J=8.0 Hz), 8.18 (1H, d, J=8.5Hz), 8.29 (2H, s), 9.08 (1H, d, J=2.2 Hz).

ESI-MS m/z 454[M+H]⁺.

Example 12(S,E)-N-(4-amino-6-methylene-5-(quinolin-3-yl)-7,8-dihydro-6H-pyrimido[5,4-b]pyrrolizin-7-yl)-4-(isopropyl(methyl)amino)but-2-enamide(Compound 12)

The same synthesis as in Example 4 was performed using(E)-4-(isopropyl(methyl)amino)but-2-enoic acid hydrochloride instead ofthe (E)-4-(dimethylamino)but-2-enoic acid hydrochloride used in Example4, thereby obtaining the title compound (25.8 mg) as a yellow solid.

¹H-NMR (CDCl₃)δ: 1.00 (6H, d, J=6.6 Hz), 2.15-2.36 (3H, m), 2.79-2.88(1H, m), 3.18 (2H, dd, J=5.9, 1.5 Hz), 4.01 (1H, dd, J=11.5, 5.0 Hz),4.70 (1H, dd, J=11.5, 8.0 Hz), 5.11 (2H, s), 5.21 (1H, d, J=1.8 Hz),5.43 (1H, d, J=1.8 Hz), 5.73-5.79 (1H, m), 6.10 (1H, dt, J=15.4, 1.5Hz), 6.74 (1H, brs), 6.94 (1H, dt, J=15.4, 5.9 Hz), 7.61-7.65 (1H, m),7.77-7.81 (1H, m), 7.87 (1H, d, J=8.5 Hz), 8.16 (1H, d, J=8.8 Hz),8.23-8.26 (2H, m), 9.05 (1H, d, J=2.2 Hz).

ESI-MS m/z 468[M+H]⁺.

Example 13(R)—N-(4-amino-6-methylene-5-(quinolin-3-yl)-6,7,8,9-tetrahydropyrimido[5,4-b]indolizin-7-yl)acrylamide(Compound 13)

Step 1: Synthesis of (R)-tert-butyl(1-hydroxy-5-(methylthio)pentan-3-yl)carbamate

N-methylmorpholine (3.63 ml) and ethyl chloroformate (3.01 ml) wereadded to a solution of(R)-3-((tert-butoxycarbonyl)amino)-5-(methylthio)pentanoic acid (7.92 g)in THF (79.2 ml) at -10° C. After stirring at -10° C. for 15 minutes,the generated insoluble matter was filtered off. An aqueous solution ofsodium borohydride (1.55 g) (15 ml) was added to the filtrate at −10°C., and the mixture was stirred at −10° C. for 1 hour. A saturatedaqueous ammonium chloride solution was added to the reaction mixture,and the mixture was stirred at room temperature for 30 minutes. Ethylacetate was added to the reaction mixture to separate the organic layer.The organic layer was washed with a 0.5 N aqueous potassium hydrogensulfate solution, water, a 0.5 N aqueous sodium hydroxide solution, anda saturated sodium chloride solution, and dried over anhydrous sodiumsulfate. The solvent was then distilled off under reduced pressure. Theresulting residue was purified by silica gel column chromatography(developing solvent: chloroform/ethyl acetate), thereby obtaining thetitle compound as a light-yellow, oily substance (7.18 g).

Step 2: Synthesis of tert-butyl((3R)-1-hydroxy-5-(methylsulfinyl)pentan-3-yl)carbamate

A suspension of sodium periodate (7.0 g) in water (32 ml) was added to asolution of the (R)-tert-butyl(1-hydroxy-5-(methylthio)pentan-3-yl)carbamate (8.16 g) obtained in Step1 in methanol (98 ml) at a temperature 10° C. or lower, and the mixturewas stirred at room temperature for 2 hours. The generated insolublematter was filtered off, and the filtrate was distilled off underreduced pressure. The resulting residue was dissolved in a saturatedsodium chloride solution, followed by extraction with chloroform 3times. The organic layer was dried over anhydrous sodium sulfate, andthe solvent was distilled off under reduced pressure, thereby obtainingthe title compound (9.38 g) as a light-yellow solid.

Step 3: Synthesis of (R)-tert-butyl (5-hydroxypent-1-en-3-yl)carbamate

Sodium acetate (13.45 g) was added to a solution of the tert-butyl((3R)-1-hydroxy-5-(methylsulfinyl)pentan-3-yl)carbamate (9.38 g)obtained in Step 2 in 1,2-dichlorobenzene (140 ml) at room temperature.The mixture was stirred at an internal temperature of 166° C. for 18hours. After cooling the reaction mixture, the insoluble matter wasfiltered off, and 1,2-dichlorobenzene was distilled off under reducedpressure. The resulting residue was dissolved in ethyl acetate, washedwith a sodium hypochlorite aqueous solution, water, and a saturatedsodium chloride solution, and dried over anhydrous sodium sulfate. Thesolvent was then distilled off under reduced pressure. The resultingresidue was purified by silica gel column chromatography (developingsolvent: hexane/ethyl acetate), thereby obtaining the title compound(2.50 g) as a light-yellow, oily substance.

Step 4: Synthesis of (R)-tert-butyl(5-(4-chloro-5-iodo-7H-pyrrolo[2,3-d]pyrimidin-7-yl)pent-1-en-3-yl)carbamate

Triphenylphosphine (3.25 g) was added to and dissolved in a solution ofthe (R)-tert-butyl (5-hydroxypent-1-en-3-yl)carbamate (2.5 g) obtainedin Step 3 and 4-chloro-5-iodo-7H-pyrrolo[2,3-d]pyrimidine (2.31 g) inDME (23 ml) under ice-cooling. Thereafter, diisopropyl azodicarboxylate(2.44 ml) was gradually added. The reaction mixture was stirred underice-cooling for 30 minutes, and at room temperature for 1 hour, and thesolvent was then distilled off under reduced pressure. The resultingresidue was dissolved in ethyl acetate, washed with water, and driedover anhydrous sodium sulfate. The solvent was then distilled off underreduced pressure. The resulting residue was purified by silica gelcolumn chromatography (developing solvent: hexane/ethyl acetate),thereby obtaining the title compound (3.49 g) as a light-yellow solid.

ESI-MS m/z 463, 465 [M+H]⁺.

Step 5: Synthesis of (R)-tert-butyl(5-(4-amino-5-iodo-7H-pyrrolo[2,3-d]pyrimidin-7-yl)pent-1-en-3-yl)carbamate

28% aqueous ammonia (17.5 ml) was added to a solution of the(R)-tert-butyl(5-(4-chloro-5-iodo-7H-pyrrolo[2,3-d]pyrimidin-7-yl)pent-1-en-3-yl)carbamate(3.49 g) obtained in Step 4 in DME (17.5 ml), and the mixture wasstirred in an autoclave at an internal temperature of 105° C. for 8hours. After cooling the reaction mixture, water (70 ml) was added, andthe mixture was stirred at room temperature for 4 hours. The resultingprecipitate was collected by filtration, washed with water, and dried,thereby obtaining the title compound (3.20 g) as a light-yellow solid.

ESI-MS m/z 444 [M+H]⁺.

Step 6: Synthesis of (R)-tert-butyl(5-(4-amino-5-(quinolin-3-yl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)pent-1-en-3-yl)carbamate

A mixture of the (R)-tert-butyl(5-(4-amino-5-iodo-7H-pyrrolo[2,3-d]pyrimidin-7-yl)pent-1-en-3-yl)carbamate(3.2 g) obtained in Step 5, 3-quinolineboronic acid (1.37 g), sodiumcarbonate (843 mg), tetrakis(triphenylphosphine)palladium (250 mg), DME(32 ml) and water (32 ml) was stirred under a nitrogen atmosphere at100° C. for 6 hours. After cooling the reaction mixture, a saturatedaqueous sodium bicarbonate solution and ethyl acetate were added. Theresulting mixture was stirred at room temperature for 30 minutes. Afterfiltering off the insoluble matter, the organic layer was separated anddried over anhydrous sodium sulfate. The solvent was then distilled offunder reduced pressure. The resulting residue was purified by silica gelcolumn chromatography (developing solvent: hexane/ethyl acetate, ethylacetate/methanol), thereby obtaining the title compound (3.21 g) as alight-orange solid.

ESI-MS m/z 445 [M+H]⁺.

Step 7: Synthesis of (R)-tert-butyl(5-(4-amino-6-bromo-5-(quinolin-3-yl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)pent-1-en-3-yl)carbamate

A solution of N-bromosuccinimide (1.35 g) in THF (23 ml) was added to asolution of the (R)-tert-butyl(5-(4-amino-5-(quinolin-3-yl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)pent-1-en-3-yl)carbamate(3.21 g) obtained in Step 6 in THF (26 ml) under ice-cooling over 30minutes. The mixture was stirred under ice-cooling for 30 minutes. Afteradding a 5% aqueous sodium thiosulfate solution to the reaction mixture,the mixture was poured into a saturated aqueous sodium bicarbonatesolution, followed by extraction with ethyl acetate. The organic layerwas washed with a saturated sodium chloride solution, and dried overanhydrous sodium sulfate. The solvent was then distilled off underreduced pressure. The resulting residue was purified by silica gelcolumn chromatography (developing solvent: hexane/ethyl acetate, ethylacetate/methanol), thereby obtaining the title compound (3.15 g) as alight-brown solid.

ESI-MS m/z 523, 525 [M+H]⁺.

Step 8: Synthesis of (R)-tert-butyl(4-amino-6-methylene-5-(quinolin-3-yl)-6,7,8,9-tetrahydropyrimido[5,4-b]indolizin-7-yl)carbamate

A 4 N sodium hydroxide aqueous solution (0.454 ml) was added to asolution of the (R)-tert-butyl(5-(4-amino-6-bromo-5-(quinolin-3-yl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)pent-1-en-3-yl)carbamate(475 mg) obtained in Step 7 in THF (5 ml). Thereafter, the mixture wasdegassed under reduced pressure, followed by nitrogen purging. Aftertetrakis(triphenylphosphine)palladium (41.6 mg) was added, the mixturewas stirred overnight while being heated under reflux. After thereaction mixture was cooled to room temperature, the reaction mixturewas poured into water, and extracted with ethyl acetate. The resultingorganic layer was washed with a saturated sodium chloride solution, anddried over anhydrous sodium sulfate. After filtration and concentration,the resulting residue was purified by silica gel chromatography(developing solvent: ethyl acetate/methanol), thereby obtaining thetitle compound (293 mg) as a yellow solid.

ESI-MS m/z 443 [M+H]⁺.

Step 9: Synthesis of(R)-6-methylene-5-(quinolin-3-yl)-6,7,8,9-tetrahydropyrimido[5,4-b]indolizin-4,7-diamine

5 N hydrochloric acid (1 ml) was added to a solution of the(R)-tert-butyl(4-amino-6-methylene-5-(quinolin-3-yl)-6,7,8,9-tetrahydropyrimido[5,4-b]indolizin-7-yl)carbamate(290 mg) obtained in Step 8 in ethanol (4 ml). Thereafter, the mixturewas stirred for 6 hours at 70° C. After the reaction mixture was cooledto room temperature, the reaction mixture was adjusted to about pH 10using a 5 N sodium hydroxide aqueous solution, followed by extractionwith chloroform. The resulting organic layer was dried over anhydroussodium sulfate, followed by filtration and concentration. The resultingresidue was purified by basic silica gel chromatography (developingsolvent: chloroform/methanol), thereby obtaining the title compound (218mg) as a light-brown solid.

ESI-MS m/z 343 [M+H]⁺.

Step 10: Synthesis of Compound 13

Diisopropylethylamine (0.129 ml) and a solution of acryloyl chloride(56.1 mg) in chloroform (0.5 ml) were added to a solution of the(R)-6-methylene-5-(quinolin-3-yl)-6,7,8,9-tetrahydropyrimido[5,4-b]indolizin-4,7-diamine(215 mg) obtained in Step 9 in chloroform (4 ml) under ice-cooling, andthe resulting mixture was stirred for 30 minutes. After the reactionmixture was concentrated, the residue was purified by silica gelchromatography (developing solvent: ethyl acetate/methanol), therebyobtaining the title compound (117 mg) as a yellow solid.

¹H-NMR (CDCl₃)δ: 2.28-2.41 (2H, m), 4.32-4.50 (2H, m), 4.64 (2H, brs),4.97 (1H, s), 5.07 (1H, s), 5.05-5.12 (1H, m), 5.72 (1H, dd, J=10.2, 1.2Hz), 5.75-5.85 (1H, m), 6.14 (1H, dd, J=16.8, 10.2 Hz), 6.36 (1H, dd,J=16.8, 1.2 Hz), 7.61-7.68 (1H, m), 7.77-7.84 (1H, m), 7.87 (1H, d,J=8.4 Hz), 8.18 (1H, d, J=8.4 Hz), 8.27 (1H, d, J=2.0 Hz), 8.33 (1H, s),8.98 (1H, d, J=2.0 Hz).

ESI-MS m/z 397[M+H]⁺.

Example 14(S)—N-(4-amino-6-methyl-5-(quinolin-3-yl)-8,9-dihydropyrimido[5,4-b]indolizin-8-yl)acrylamide(Compound 14)

Step 1: Synthesis of (S)-methyl2-((tert-butoxycarbonyl)amino)pent-4-enoate

5 N sodium hydroxide aqueous solution (2.1 ml) and di-tert-butyldicarbonate (2.128 ml) were added to a suspension of(S)-2-amino-4-pentenoic acid (1.016 g) in methanol (20 ml) at roomtemperature, and the resulting mixture was stirred for 8 hours at thesame temperature. Di-tert-butyl dicarbonate (0.304 ml) was added at roomtemperature, and the mixture was stirred for 1 hour at the sametemperature. 4-hydroxy-1H-benzotriazole (1.796 g) and1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (3.882 g)were added to the reaction mixture at room temperature, and the mixturewas stirred overnight at the same temperature. The reaction mixture wasconcentrated under reduced pressure, and then was poured into asaturated aqueous sodium bicarbonate solution, followed by extractionwith ethyl acetate. The resulting organic layer was dried over anhydroussodium sulfate, followed by filtration and concentration. The resultingresidue was purified by silica gel chromatography (developing solvent:chloroform/methanol), thereby obtaining the title compound (2.0311 g) asa light-yellow, oily substance.

Step 2: Synthesis of (S)-tert-butyl (1-hydroxypent-4-en-2-yl)carbamate

Lithium aluminium hydride (668.2 mg) was added to a solution of the(S)-methyl2-((tert-butoxycarbonyl)amino)pent-4-enoate (1.983 g) obtainedin Step 1 in THF (50 ml) under ice cooling, and the mixture was stirredfor 1.5 hours at the same temperature. Sodium sulfate decahydrate(1.1375 g) and THF (10 ml) were added to the reaction mixture at roomtemperature, and the mixture was stirred overnight at the sametemperature. The insoluble matter was filtered off. After washing withTHF and ethyl acetate, the filtrate was concentrated under reducedpressure. The resulting residue was purified by silica gelchromatography (developing solvent: hexane/ethyl acetate), therebyobtaining the title compound (1.2601 g) as a light-yellow, oilysubstance.

Step 3: Synthesis of (S)-tert-butyl(1-(4-chloro-5-iodo-7H-pyrrolo[2,3-d]pyrimidin-7-yl)pent-4-en-2-yl)carbamate

The same synthesis as in Step 4 of Example 13 was performed using the(S)-tert-butyl (1-hydroxypent-4-en-2-yl)carbamate obtained in Step 2instead of the (R)-tert-butyl (5-hydroxypent-1-en-3-yl)carbamate used inStep 4 of Example 13, thereby obtaining the title compound (2.7679 g) asa light-yellow solid.

ESI-MS m/z 463, 465 [M+H]⁺.

Step 4: Synthesis of (S)-tert-butyl(1-(4-amino-6-bromo-5-(quinolin-3-yl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)pent-4-en-2-yl)carbamate

The same synthesis as in Steps 5 to 7 of Example 13 was performed usingthe (S)-tert-butyl(1-(4-chloro-5-iodo-7H-pyrrolo[2,3-d]pyrimidin-7-yl)pent-4-en-2-yl)carbamateobtained in Step 3 instead of the (R)-tert-butyl(5-(4-chloro-5-iodo-7H-pyrrolo[2,3-d]pyrimidin-7-yl)pent-1-en-3-yl)carbamateused in Step 5 of Example 13, thereby obtaining the title compound(2.669 g) as a light-brown solid.

ESI-MS m/z 523, 525 [M+H]⁺.

Step 5: Synthesis of (S)-tert-butyl(4-amino-6-methyl-5-(quinolin-3-yl)-8,9-dihydropyrimido[5,4-b]indolizin-8-yl)carbamateand (S)-tert-butyl(4-amino-6-methylene-5-(quinolin-3-yl)-6,7,8,9-tetrahydropyrimido[5,4-b]indolizin-8-yl)carbamate

A 4 N sodium hydroxide aqueous solution (5.1 ml) andtetrakis(triphenylphosphine)palladium (235.4 mg) were added to asolution of the (S)-tert-butyl(1-(4-amino-6-bromo-5-(quinolin-3-yl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)pent-4-en-2-yl)carbamate(2.669 g) obtained in Step 4 in THF (50 ml), and the mixture wasdegassed under reduced pressure, followed by nitrogen purging. Thereaction mixture was stirred overnight while being heated under reflux.After the reaction mixture was cooled to room temperature, the reactionmixture was poured into a saturated aqueous sodium bicarbonate solution,and extracted with ethyl acetate. The resulting organic layer was driedover anhydrous sodium sulfate. After filtration and concentration, theresulting residue was purified by silica gel chromatography (developingsolvent: chloroform/methanol), thereby obtaining a mixture of the titlecompound (2.427 g) as a yellow solid.

ESI-MS m/z 443 [M+H]⁺.

Step 6: Synthesis of(S)-6-methyl-5-(quinolin-3-yl)-8,9-dihydropyrimido[5,4-b]indolizin-4,8-diamineand(S)-6-methylene-5-(quinolin-3-yl)-6,7,8,9-tetrahydropyrimido[5,4-b]indolizin-4,8-diamine

5 N hydrochloric acid (5.1 ml) was added to a solution of a mixture(2.427 g) of the (S)-tert-butyl(4-amino-6-methyl-5-(quinolin-3-yl)-8,9-dihydropyrimido[5,4-b]indolizin-8-yl)carbamateand (S)-tert-butyl(4-amino-6-methylene-5-(quinolin-3-yl)-6,7,8,9-tetrahydropyrimido[5,4-b]indolizin-8-yl)carbamateobtained in Step 5 in ethanol (20 ml), and the mixture was stirredovernight at 60° C. After an additional 5 N hydrochloric acid (5.1 ml)was added, the mixture was stirred for 10 hours while being heated underreflux. After the reaction mixture was cooled to room temperature, thereaction mixture was diluted with water, and washed with chloroform. A 5N sodium hydroxide aqueous solution (10 ml) was added to the aqueouslayer, followed by extraction with chloroform. The resulting organiclayer was dried over anhydrous sodium sulfate, followed by filtrationand concentration. The resulting residue was purified by basic silicagel chromatography (developing solvent: ethyl acetate/methanol), therebyobtaining a mixture of the title compound (1.4098 g) as a light-yellowsolid.

ESI-MS m/z 343 [M+H]⁺.

Step 7: Synthesis of Compound 14

Diisopropylethylamine (0.8452 ml) and a solution of acryloyl chloride(0.35 ml) in acetonitrile (3.5 ml) were successively added to a solutionof a mixture (1.407 g) of the(S)-6-methyl-5-(quinolin-3-yl)-8,9-dihydropyrimido[5,4-b]indolizin-4,8-diamineand(S)-6-methylene-5-(quinolin-3-yl)-6,7,8,9-tetrahydropyrimido[5,4-b]indolizin-4,8-diamineobtained in Step 6 in acetonitrile (10 ml) and water (10 ml) at 0° C.,and the mixture was stirred for 45 minutes at the same temperature.After the reaction mixture was diluted with water, the reaction mixturewas poured into a saturated aqueous sodium bicarbonate solution,followed by extraction with ethyl acetate. After the organic layer wasdried over anhydrous sodium sulfate, the solvent was distilled off underreduced pressure. The resulting residue was purified by silica gelcolumn chromatography (developing solvent: ethyl acetate/methanol),thereby obtaining the title compound (897.1 mg) as a light-yellow solid.

¹H-NMR (DMSO-d₆)δ: 1.51 (3H, s), 4.17 (1H, dd, J=13.2, 5.1 Hz), 4.25(1H, dd, J=13.2, 5.1 Hz), 4.73-4.83 (1H, m), 5.61 (1H, dd, J=9.8, 2.7Hz), 5.65-6.00 (2H, brs), 5.81 (1H, dd, J=5.1, 1.2 Hz), 6.14 (1H, dd,J=17.1, 2.4 Hz), 6.24 (1H, dd, J=17.1, 9.8 Hz), 7.67 (1H, ddd, J=8.1,7.1, 1.0 Hz), 7.81 (1H, ddd, J=8.3, 7.1, 1.5 Hz), 8.05 (1H, d, J=8.1Hz), 8.09 (1H, d, J=8.3 Hz), 8.13 (1H, s), 8.39 (1H, brs), 8.46 (1H, d,J=7.3 Hz), 8.94 (1H, s).

ESI-MS m/z 397[M+H]⁺.

Example 15(S)—N-(4-amino-6-methylene-5-(quinolin-3-yl)-6,7,8,9-tetrahydropyrimido[5,4-b]indolizin-8-yl)acrylamide(Compound 15)

The mixed fraction obtained by the silica gel column chromatographypurification in Step 7 of Example 14 was purified by reversed-phasepreparative HPLC, thereby obtaining the title compound (32.2 mg) as alight-yellow solid.

¹H-NMR (DMSO-d₆)δ: 2.56-2.70 (1H, m), 2.72-2.82 (1H, m), 4.19 (1H, dd,J=14.1, 8.3 Hz), 4.25-4.36 (1H, m), 4.72 (1H, d, J=13.7 Hz), 5.62 (1H,dd, J=10.0, 2.2 Hz), 5.80 (1H, dt, J=12.2, 4.6 Hz), 6.00-6.20 (3H, m),6.20-6.31 (2H, m), 7.65 (1H, t, J=8.1 Hz), 7.80 (1H, t, 8.3 Hz), 8.05(1H, d, J=8.5 Hz), 8.08 (1H, d, J=8.3 Hz), 8.15 (1H, s), 8.28-8.39 (2H,m), 8.87 (1H, d, J=2.2 Hz).

ESI-MS m/z 397[M+H]⁺.

Example 16(R)—N-(4-amino-6-methyl-5-(quinolin-3-yl)-8,9-dihydropyrimido[5,4-b]indolizin-8-yl)acrylamide(Compound 16)

Step 1: Synthesis of (R)-tert-butyl(1-(4-chloro-5-iodo-7H-pyrrolo[2,3-d]pyrimidin-7-yl)pent-4-en-2-yl)carbamate

The same synthesis as in Steps 1 to 3 of Example 14 was performed using(R)-2-amino-4-pentenoic acid instead of the (S)-2-amino-4-pentenoic acidused in Step 1 of Example 14, thereby obtaining the title compound(1.4217 g) as a light-yellow solid.

ESI-MS m/z 463, 465 [M+H]⁺.

Step 2: Synthesis of Compound 16

The same synthesis as in Steps 4 to 7 of Example 14 was performed usingthe (R)-tert-butyl(1-(4-chloro-5-iodo-7H-pyrrolo[2,3-d]pyrimidin-7-yl)pent-4-en-2-yl)carbamateobtained in Step 1, thereby obtaining the title compound (21.0 mg) as alight-yellow solid.

¹H-NMR (DMSO-d₆)δ: 1.51 (3H, s), 4.17 (1H, dd, J=13.2, 5.1 Hz), 4.25(1H, dd, J=13.2, 5.1 Hz), 4.73-4.83 (1H, m), 5.61 (1H, dd, J=9.8, 2.7Hz), 5.65-6.00 (2H, brs), 5.81 (1H, dd, J=5.1, 1.2 Hz), 6.14 (1H, dd,J=17.1, 2.4 Hz), 6.24 (1H, dd, J=17.1, 9.8 Hz), 7.67 (1H, t, J=7.1 Hz),7.81 (1H, t, J=7.6 Hz), 8.05 (1H, d, J=8.1 Hz), 8.09 (1H, d, J=8.6 Hz),8.13 (1H, s), 8.39 (1H, brs), 8.46 (1H, d, J=7.1 Hz), 8.94 (1H, s).

ESI-MS m/z 397[M+H]⁺.

Example 17(R)—N-(4-amino-6-methylene-5-(quinolin-3-yl)-6,7,8,9-tetrahydropyrimido[5,4-b]indolizin-8-yl)acrylamide(Compound 17)

The mixed fraction obtained by the silica gel column chromatographypurification in Example 16 was purified in the same method as in Example15, thereby obtaining the title compound (8.4 mg) as a light-yellowsolid.

¹H-NMR (DMSO-d₆)δ: 2.57-2.70 (1H, m), 2.71-2.82 (1H, m), 4.19 (1H, dd,J=13.2, 8.3 Hz), 4.25-4.36 (1H, m), 4.72 (1H, d, J=13.2 Hz), 5.62 (1H,dd, J=10.0, 2.2 Hz), 5.80 (1H, dt, J=12.2, 4.6 Hz), 6.00-6.20 (3H, m),6.20-6.31 (2H, m), 7.65 (1H, t, J=7.6 Hz), 7.79 (1H, ddd, J=8.3, 7.1,1.5 Hz), 8.05 (1H, d, J=8.5 Hz), 8.08 (1H, d, J=8.5 Hz), 8.15 (1H, s),8.33 (1H, d, J=2.2 Hz), 8.36 (1H, d, J=7.6 Hz), 8.87 (1H, d, J=2.2 Hz).

ESI-MS m/z 397[M+H]⁺.

Example 18(S)—N-(4-amino-6-methylene-5-(quinolin-3-yl)-7,8,9,10-tetrahydro-6H-pyrimido[5′,4′:4,5]pyrrolo[1,2-a]azepin-8-yl)acrylamide(Compound 18)

Step 1: Synthesis of (S)-tert-butyl (1-hydroxyhex-5-en-3-yl)carbamate

N-methylmorpholine (2.1 g) was added to a solution of(S)-3-((tert-butoxycarbonyl)amino)hex-5-enoic acid (4.0 g) in THF (40ml), and ethyl chloroformate (1.75 ml) was gradually added at −10° C.Thereafter, the mixture was stirred for 20 minutes at the sametemperature, and the generated insoluble matter was filtered throughCelite. A solution of sodium tetrahydroborate (904 mg) in water (8 ml)was gradually added to the resulting filtrate at −10° C. After themixture was stirred for 30 minutes at the same temperature, a saturatedaqueous ammonium chloride solution was added. After extraction withethyl acetate, washing was performed using a 0.5 N potassium hydrogensulfate aqueous solution, a saturated aqueous sodium bicarbonatesolution, and a saturated sodium chloride solution in this order. Theresulting organic layer was dried over anhydrous sodium sulfate and thenfiltered, followed by concentration. The resulting residue was purifiedby silica gel chromatography (developing solvent: hexane/ethyl acetate),thereby obtaining the title compound (3.54 g) as an oily substance.

Step 2: Synthesis of Compound 18

The same synthesis as in Steps 4 to 10 of Example 13 was performed usingthe (S)-tert-butyl (1-hydroxyhex-5-en-3-yl)carbamate obtained in Step 1instead of the (R)-tert-butyl (5-hydroxypent-1-en-3-yl)carbamate used inStep 4 of Example 13, thereby obtaining the title compound (147 mg) as acolorless solid.

¹H-NMR (DMSO-d₆)δ: 1.68-1.82 (1H, m), 2.17-2.25 (1H, m), 2.42-2.53 (1H,m), 2.75-2.83 (1H, m), 3.86-4.16 (2H, m), 4.64-4.78 (1H, m), 4.84 (1H,d, J=1.7 Hz), 5.26 (1H, s), 5.60 (1H, dd, J=10.0, 2.4 Hz), 6.00 (2H,brs), 6.11 (1H, dd, J=17.1, 2.4 Hz), 6.27 (1H, dd, J=17.1, 10.0 Hz),7.60-7.64 (1H, m), 7.73-7.77 (1H, m), 7.99-8.04 (2H, m), 8.17 (1H, s),8.22 (1H, d, J=8.0 Hz), 8.30 (1H, d, J=2.0 Hz), 8.78 (1H, d, J=2.0 Hz).

ESI-MS m/z 411[M+H]⁺.

Example 19(S,E)-N-(4-amino-6-ethylidene-5-(quinolin-3-yl)-6,7,8,9-tetrahydropyrimido[5,4-b]indolizin-8-yl)acrylamide(Compound 19)

Step 1: Synthesis of (S)-methyl3-((tert-butoxycarbonyl)amino)-4-hydroxybutanoate

N-methylmorpholine (9.78 ml) and ethyl chloroformate (8.09 ml) wereadded to a solution of(S)-2-((tert-butoxycarbonyl)amino)-4-methoxy-4-oxobutanoic acid (20.0 g)in THF (200 ml) under ice-cooling, and the mixture was stirred for 1hour under ice-cooling. The generated insoluble matter was filtered off.A solution of sodium borohydride (4.14 g) in water (41.4 ml) was addedto the filtrate under ice-cooling, and the mixture was stirred for 30minutes under ice-cooling. A 0.5 N potassium hydrogen sulfate aqueoussolution and ethyl acetate were added to the reaction mixture. Theorganic layer was separated, washed with a saturated aqueous sodiumbicarbonate solution and a saturated sodium chloride solution, and driedover anhydrous magnesium sulfate. The solvent was then distilled offunder reduced pressure, thereby obtaining the title compound (10.82 g).

Step 2: Synthesis of (S)-tert-butyl4-(2-methoxy-2-oxoethyl)-2,2-dimethyloxazolidine-3-carboxylate

2,2-dimethoxypropane (28.52 ml) and boron trifluoride-diethylethercomplex (0.294 ml) were added to a solution of the (S)-methyl3-((tert-butoxycarbonyl)amino)-4-hydroxybutanoate (10.82 g) obtained inStep 1 in acetone (108.2 ml) at room temperature, and the mixture wasstirred for 4 hours at the same temperature. The reaction mixture waspoured into a saturated aqueous sodium bicarbonate solution, andextracted with ethyl acetate. The resulting organic layer was dried overanhydrous magnesium sulfate, and then filtered, followed byconcentration. The resulting residue was purified by silica gelchromatography (developing solvent: hexane/ethyl acetate), therebyobtaining the title compound (8.72 g).

Step 3: Synthesis of (S)-tert-butyl4-(2-hydroxyethyl)-2,2-dimethyloxazolidine-3-carboxylate

A 1 M diisobutylaluminium hydride-toluene solution (65.7 ml) was addedto a solution of the (S)-tert-butyl4-(2-methoxy-2-oxoethyl)-2,2-dimethyloxazolidine-3-carboxylate (8.71 g)obtained in Step 2 in methylene chloride (87.1 ml) under ice cooling,and the mixture was stirred for 2 hours under ice cooling. A 5%potassium sodium tartrate aqueous solution and ethyl acetate were addedto the reaction mixture, and the mixture was stirred overnight at roomtemperature. The resulting organic layer was dried over anhydrousmagnesium sulfate, and then filtered, followed by concentration. Theresulting residue was purified by silica gel chromatography (developingsolvent: hexane/ethyl acetate), thereby obtaining the title compound(5.94 g).

Step 4: Synthesis of (S)-tert-butyl2,2-dimethyl-4-(2-oxoethyl)oxazolidine-3-carboxylate

Triethylamine (16.9 ml) and a sulfur trioxide pyridine complex (12.56 g)were added to a solution of the (S)-tert-butyl4-(2-hydroxyethyl)-2,2-dimethyloxazolidine-3-carboxylate (5.94 g)obtained in Step 3 in DMSO (59.4 ml) at room temperature, and themixture was stirred for 1 hour at the same temperature. The reactionmixture was poured into a saturated aqueous sodium bicarbonate solution,and extracted with ethyl acetate. The resulting organic layer was driedover anhydrous magnesium sulfate, and then filtered, followed byconcentration. The resulting residue was purified by silica gelchromatography (developing solvent: hexane/ethyl acetate), therebyobtaining the title compound (6.06 g).

Step 5: Synthesis of (S)-tert-butyl4-(but-2-en-1-yl)-2,2-dimethyloxazolidine-3-carboxylate

A 2.69 M n-butyllithium-hexane solution (7.0 ml) was added to asuspension of ethyltriphenylphosphonium bromide (5.60 g) in THF (25.2ml) under ice-cooling, and the mixture was stirred for 30 minutes underice-cooling. A solution of the (S)-tert-butyl2,2-dimethyl-4-(2-oxoethyl)oxazolidine-3-carboxylate (3.06 g) obtainedin Step 4 in THF (3.06 ml) was added to the reaction mixture underice-cooling, and the mixture was stirred for 14 hours at roomtemperature. Hexane was added to the reaction mixture, and the insolublematter was filtered off, followed by washing with THF−hexane=2/1. Thefiltrate was concentrated under reduced pressure. The resulting residuewas purified by silica gel chromatography (developing solvent:hexane/ethyl acetate), thereby obtaining the title compound (1.81 g).

Step 6: Synthesis of (S)-tert-butyl (1-hydroxyhex-4-en-2-yl)carbamate

A 4 N hydrogen chloride-dioxane solution (18.1 ml) was added to the(S)-tert-butyl 4-(but-2-en-1-yl)-2,2-dimethyloxazolidine-3-carboxylate(1.81 g) obtained in Step 5 at room temperature, and the mixture wasstirred for 2 hours at 70° C. The reaction mixture was cooled, and thenconcentrated under reduced pressure. The resulting residue was dissolvedin THF (18.1 ml), and a saturated aqueous sodium bicarbonate solution(18.1 ml) and di-tert-butyl dicarbonate (1.53 g) were added thereto atroom temperature. The mixture was stirred overnight at the sametemperature. The reaction mixture was poured into water, and extractedwith ethyl acetate. The resulting organic layer was dried over anhydrousmagnesium sulfate, and then filtered, followed by concentration. Theresulting residue was purified by silica gel chromatography (developingsolvent: hexane/ethyl acetate), thereby obtaining the title compound(1.57 g).

Step 7: Synthesis of Compound 19

The same synthesis as in Steps 4 to 10 of Example 13 was performed usingthe (S)-tert-butyl (1-hydroxyhex-4-en-2-yl)carbamate obtained in Step 6instead of the (R)-tert-butyl (5-hydroxypent-1-en-3-yl)carbamate used inStep 4 of Example 13, thereby obtaining the title compound (327 mg).

¹H-NMR (DMSO-d₆)δ: 1.47 (3H, d, J=7.1 Hz), 2.64-2.70 (1H, m), 2.81-2.85(1H, m), 4.00-4.09 (1H, m), 4.32-4.43 (2H, m), 5.41-5.46 (1H, m),5.60-5.63 (1H, m), 5.82 (2H, brs), 6.11-6.16 (1H, m), 6.21-6.31 (1H, m),8.05 (1H, d, J=7.3 Hz), 8.08 (1H, d, J=8.5 Hz), 8.13 (1H, s), 8.35-8.39(2H, m), 8.87-8.92 (1H, m).

ESI-MS m/z 411[M+H]⁺.

Example 20 Mixture of(S)—N-(4-amino-6-isopropyl-5-(quinolin-3-yl)-8,9-dihydropyrimido[5,4-b]indolizin-8-yl)acrylamide(Compound 20A) and(S)—N-(4-amino-6-(propan-2-ylidene)-5-(quinolin-3-yl)-6,7,8,9-tetrahydropyrimido[5,4-b]indolizin-8-yl)acrylamide(Compound 20B)

The same synthesis as in Example 19 was performed usingisopropyltriphenylphosphonium iodide instead of theethyltriphenylphosphonium bromide used in Step 5 of Example 19, therebyobtaining a mixture (18.9 mg) of the title compound.

¹H-NMR (DMSO-d₆)δ: 0.85-4.71 (11H, m), 5.63-5.68 (1H, m), 5.88 (1H,brs), 6.03 (1H, brs), 6.12-6.19 (1H, m), 6.24-6.31 (1H, m), 7.59-7.64(1H, m), 7.72-7.78 (1H, m), 7.96-7.99 (1H, m), 8.01-8.05 (1H, m), 8.16(1H, d, J=3.4 Hz), 8.24-8.28 (1H, m), 8.39-8.44 (1H, m), 8.90 (1H, d,J=2.0 Hz).

ESI-MS m/z 425[M+H]⁺.

Example 21(R)—N-(4-amino-6-methylene-5-(quinolin-3-yl)-7,8-dihydro-6H-pyrimido[5,4-b]pyrrolizin-7-yl)-N-methylacrylamide(Compound 21)

Step 1: Synthesis of (R)-tert-butyl(1-(4-chloro-5-iodo-7H-pyrrolo[2,3-d]pyrimidin-7-yl)but-3-en-2-yl)carbamate

The same synthesis as in Step 1 of Example 1 was performed using(R)-tert-butyl (1-hydroxybut-3-en-2-yl)carbamate instead of the(S)-tert-butyl (1-hydroxybut-3-en-2-yl)carbamate used in Step 1 ofExample 1, thereby obtaining the title compound (1.083 g) as alight-yellow solid.

ESI-MS m/z 448, 450 [M+H]⁺.

Step 2: Synthesis of (R)-tert-butyl(1-(4-chloro-5-iodo-7H-pyrrolo[2,3-d]pyrimidin-7-yl)but-3-en-2-yl)(methyl)carbamate

Methyl iodide (1.58 ml), and sodium hydride (224 mg) dispersed in liquidparaffin were added to a solution of the (R)-tert-butyl(1-(4-chloro-5-iodo-7H-pyrrolo[2,3-d]pyrimidin-7-yl)but-3-en-2-yl)carbamate(2.28g) obtained in Step 1 in DMF (11.4 ml) at room temperature, and themixture was stirred for 1 hour at the same temperature. The resultingmixture was poured into water, and extracted with ethyl acetate. Theresulting organic layer was washed with water and a saturated sodiumchloride solution, and was dried over anhydrous sodium sulfate;thereafter, the solvent was distilled off under reduced pressure. Theresulting residue was purified by silica gel column chromatography(developing solvent: hexane/ethyl acetate), thereby obtaining the titlecompound (2.41 g) as a light-yellow solid.

ESI-MS m/z 462, 464 [M+H]⁺.

Step 3: Synthesis of Compound 21

The same synthesis as in Steps 5 to 10 of Example 13 was performed usingthe (R)-tert-butyl(1-(4-chloro-5-iodo-7H-pyrrolo[2,3-d]pyrimidin-7-yl)but-3-en-2-yl)(methyl)carbamateobtained in Step 2 instead of the (R)-tert-butyl(5-(4-chloro-5-iodo-7H-pyrrolo[2,3-d]pyrimidin-7-yl)pent-1-en-3-yl)carbamateused in Step 5 of Example 13, thereby obtaining the title compound (256mg) as a light-yellow solid.

¹H-NMR (DMSO-d₆)δ: 2.75 and 2.94 (total 3H, each s), 4.06-4.20 (1H, m),4.46-4.72 (1H, m), 4.81-4.89 (1H, m), 5.26-5.35 (1H, m), 5.70-5.80 (1H,m), 5.98-6.37 (4H, m), 6.77-7.01 (1H, m), 7.62-7.68 (1H, m), 7.77-7.83(1H, m), 8.03-8.10 (2H, m), 8.14-8.18 (1H, m), 8.41-8.45 (1H, m),8.96-9.01 (1H, m).

ESI-MS m/z 397[M+H]⁺.

Example 22(R)—N-(4-amino-6-methylene-5-(quinolin-3-yl)-6,7,8,9-tetrahydropyrimido[5,4-b]indolizin-8-yl)-N-methylacrylamide(Compound 22)

Step 1: Synthesis of (R)-tert-butyl(1-(4-chloro-5-iodo-7H-pyrrolo[2,3-d]pyrimidin-7-yl)pent-4-en-2-yl)(methyl)carbamate

The same synthesis as in Step 2 of Example 21 was performed using the(R)-tert-butyl(1-(4-chloro-5-iodo-7H-pyrrolo[2,3-d]pyrimidin-7-yl)pent-4-en-2-yl)carbamateobtained in Step 1 of Examples 16 and 17 instead of the (R)-tert-butyl(1-(4-chloro-5-iodo-7H-pyrrolo[2,3-d]pyrimidin-7-yl)but-3-en-2-yl)carbamateused in Step 2 of Example 21, thereby obtaining the title compound(2.743 g) as a light-yellow solid.

ESI-MS m/z 477, 479 [M+H]⁺.

Step 2: Synthesis of (R)-tert-butyl(4-amino-6-methylene-5-(quinolin-3-yl)-6,7,8,9-tetrahydropyrimido[5,4-b]indolizin-8-yl)(methyl)carbamate

The same synthesis as in Steps 5 to 8 of Example 13 was performed usingthe (R)-tert-butyl(1-(4-chloro-5-iodo-7H-pyrrolo[2,3-d]pyrimidin-7-yl)pent-4-en-2-yl)(methyl)carbamateobtained in Step 1 instead of the (R)-tert-butyl(5-(4-chloro-5-iodo-7H-pyrrolo[2,3-d]pyrimidin-7-yl)pent-1-en-3-yl)carbamateused in Step 5 of Example 13, thereby obtaining the title compound(1.366 g) as a light-brown solid.

ESI-MS m/z 457 [M+H]⁺.

Step 3: Synthesis of(R)—N⁸-methyl-6-methylene-5-(quinolin-3-yl)-6,7,8,9-tetrahydropyrimido[5,4-b]indolizin-4,8-diamine

5 N hydrochloric acid (0.5 ml) was added to a solution of the(R)-tert-butyl(4-amino-6-methylene-5-(quinolin-3-yl)-6,7,8,9-tetrahydropyrimido[5,4-b]indolizin-8-yl)(methyl)carbamate(228 mg) obtained in Step 2 in ethanol (3 ml), and the mixture wasstirred for 4 days at 50° C. After cooling, the reaction mixture wasbasified with a 5 N aqueous sodium hydroxide solution, followed byextraction with chloroform. After the resulting organic layer was driedover anhydrous sodium sulfate, the solvent was distilled off underreduced pressure. The resulting residue was purified by basic silica gelcolumn chromatography (developing solvent: chloroform/methanol), therebyobtaining the title compound (197 mg) as a light-brown solid.

ESI-MS m/z 357 [M+H]⁺.

Step 4: Synthesis of Compound 22

The same synthesis as in Step 10 of Example 13 was performed using the(R)—N⁸-methyl-6-methylene-5-(quinolin-3-yl)-6,7,8,9-tetrahydropyrimido[5,4-b]indolizin-4,8-diamineobtained in Step 3 instead of the((R)-6-methylene-5-(quinolin-3-yl)-6,7,8,9-tetrahydropyrimido[5,4-b]indolizin-4,7-diamineused in Step 10 of Example 13, thereby obtaining the title compound(68.5 mg) as a light-yellow solid.

¹H-NMR (DMSO-d₆)δ: 2.53-2.68 (1H, m), 2.88-3.11 (4H, m), 4.04-4.20 (1H,m), 4.34-4.96 (4H, m), 5.58-6.21 (4H, m), 6.72-7.01 (1H, m), 7.62-7.68(1H, m), 7.78-7.83 (1H, m), 8.02-8.10 (2H, m), 8.15 (1H, brs), 8.37-8.44(1H, m), 8.86-8.90 (1H, m).

ESI-MS m/z 411[M+H]⁺.

Example 23(R)—N-(4-amino-6-methyl-5-(quinolin-3-yl)-8,9-dihydropyrimido[5,4-b]indolizin-8-yl)-N-methylacrylamide(Compound 23)

Step 1: Synthesis of(R)—N⁸,6-dimethyl-5-(quinolin-3-yl)-8,9-dihydropyrimido[5,4-b]indolizin-4,8-diamine

5 N hydrochloric acid (1 ml) was added to a solution of the(R)-tert-butyl(4-amino-6-methylene-5-(quinolin-3-yl)-6,7,8,9-tetrahydropyrimido[5,4-b]indolizin-8-yl)(methyl)carbamate(228 mg) obtained in Step 2 of Example 22 in ethanol (4 ml), and themixture was stirred for 24 hours while being heated under reflux. Aftercooling, the reaction mixture was basified with a 5 N aqueous sodiumhydroxide solution, followed by extraction with chloroform. After theresulting organic layer was dried over anhydrous sodium sulfate, thesolvent was distilled off under reduced pressure. The resulting residuewas purified by basic silica gel column chromatography (developingsolvent: chloroform/methanol), thereby obtaining the title compound(190.8 mg) as a light-brown solid.

ESI-MS m/z 357 [M+H]⁺.

Step 2: Synthesis of Compound 23

The same synthesis as in Step 10 of Example 13 was performed using the(R)—N⁸,6-dimethyl-5-(quinolin-3-yl)-8,9-dihydropyrimido[5,4-b]indolizin-4,8-diamineobtained in Step 1 instead of the((R)-6-methylene-5-(quinolin-3-yl)-6,7,8,9-tetrahydropyrimido[5,4-b]indolizin-4,7-diamineused in Step 10 of Example 13, thereby obtaining the title compound(150.9 mg) as a yellow solid.

¹H-NMR (DMSO-d₆)δ: 1.52 (3H, s), 2.74 and 2.88 (total 3H, each brs),4.14-4.42 (2H, m), 5.21-5.98 (5H, m), 6.09-6.22 (1H, m), 6.71-7.03 (1H,m), 7.64-7.69 (1H, m), 7.79-7.85 (1H, m), 8.02-8.07 (1H, m), 8.08-8.12(1H, m), 8.14 (1H, s), 8.43-8.49 (1H, m), 8.89-8.96 (1H, m).

ESI-MS m/z 411[M+H]⁺.

Example 24N-((7S)-4-amino-6-methyl-5-(quinolin-3-yl)-7,8-dihydro-6H-pyrimido[5,4-b]pyrrolizin-7-yl)acrylamide(Compound 24)

Step 1: Synthesis of tert-butyl((75)-4-amino-6-methyl-5-(quinolin-3-yl)-7,8-dihydro-6H-pyrimido[5,4-b]pyrrolizin-7-yl)carbamate

10% palladium-carbon (50% wet, 15.0 mg) was added to a solution of the(S)-tert-butyl(4-amino-6-methylene-5-(quinolin-3-yl)-7,8-dihydro-6H-pyrimido[5,4-b]pyrrolizin-7-yl)carbamate(15.0 mg) obtained in Step 5 of Example 1 in ethyl acetate (2ml)-ethanol (1 ml), and the mixture was stirred for 12 hours at roomtemperature under a hydrogen atmosphere. The reaction mixture wasfiltered through Celite, and the filtrate was concentrated under reducedpressure. The resulting residue was purified by preparative thin-layerchromatography (developing solvent: chloroform/methanol), therebyobtaining the title compound (10.0 mg).

ESI-MS m/z 357 [M+H]⁺.

Step 2: Synthesis of Compound 24

The same synthesis as in Steps 9 and 10 of Example 13 was performedusing the tert-butyl((7S)-4-amino-6-methyl-5-(quinolin-3-yl)-7,8-dihydro-6H-pyrimido[5,4-b]pyrrolizin-7-yl)carbamateobtained in Step 1 instead of the (R)-tert-butyl(4-amino-6-methylene-5-(quinolin-3-yl)-6,7,8,9-tetrahydropyrimido[5,4-b]indolizin-7-yl)carbamateused in Step 9 of Example 13, thereby obtaining the title compound (3.10mg) as a colorless solid.

¹H-NMR (DMSO-d₆)δ: 0.86 (2.7H, d, J=7.6 Hz), 1.08 (0.3H, d, J=7.6 Hz),3.43-4.02 (2H, m), 4.34-4.62 (1H, m), 5.08-5.19 (1H, m), 5.59-5.68 (1H,m), 5.99-6.17 (3H, m), 6.18-6.33 (1H, m), 7.62 (1H, dd, J=7.6, 7.6 Hz),7.75 (1H, dd, J=7.6, 7.6 Hz), 7.99-8.07 (2H, m), 8.12 (1H, s), 8.29-8.36(1H, m), 8.53-8.76 (1H, m), 8.98 (1H, d, J=2.2 Hz).

ESI-MS m/z 385[M+H]⁺.

Example 25(R)—N-(4-amino-6-methylene-5-(quinolin-3-yl)-7,8-dihydro-6H-pyrimido[5,4-b]pyrrolizin-7-yl)acrylamide(Compound 25)

The same synthesis as in Steps 2 to 6 of Example 1 was performed usingthe (R)-tert-butyl(1-(4-chloro-5-iodo-7H-pyrrolo[2,3-d]pyrimidin-7-yl)but-3-en-2-yl)carbamateobtained in Step 1 of Example 21 instead of the (S)-tert-butyl(1-(4-chloro-5-iodo-7H-pyrrolo[2,3-d]pyrimidin-7-yl)but-3-en-2-yl)carbamateused in Step 2 of Example 1, thereby obtaining the title compound (44.6mg) as a light-yellow solid.

¹H-NMR (DMSO-d₆)δ: 3.88-3.93 (1H, m), 4.57-4.63 (1H, m), 5.03 (1H, d,J=2.4 Hz), 5.24 (1H, d, J=2.4 Hz), 5.55-5.62 (1H, m), 5.68 (1H, dd,J=10.0, 2.4 Hz), 6.12-6.38 (4H, m), 7.65 (1H, dd, J=7.8, 7.8 Hz),7.77-7.83 (1H, m), 8.04-8.11 (2H, m), 8.15 (1H, s), 8.41 (1H, d, J=2.2Hz), 8.82 (1H, d, J=7.8 Hz), 8.98 (1H, d, J=2.2 Hz).

ESI-MS m/z 383[M+H]⁺.

Example 26(S)—N-(4-amino-6-methylene-5-(quinolin-3-yl)-7,8-dihydro-6H-pyrimido[5,4-b]pyrrolizin-7-yl)-N-methylacrylamide(Compound 26)

The same synthesis as in Steps 2 and 3 of Example 21 was performed usingthe (S)-tert-butyl(1-(4-chloro-5-iodo-7H-pyrrolo[2,3-d]pyrimidin-7-yl)but-3-en-2-yl)carbamateobtained in Step 1 of Example 1 instead of the (R)-tert-butyl(1-(4-chloro-5-iodo-7H-pyrrolo[2,3-d]pyrimidin-7-yl)but-3-en-2-yl)carbamateused in Step 2 of Example 21, thereby obtaining the title compound (143mg) as a light-yellow solid.

¹H-NMR (DMSO-d₆)δ: 2.75 and 2.94 (total 3H, each s), 4.06-4.20 (1H, m),4.46-4.72 (1H, m), 4.81-4.89 (1H, m), 5.26-5.35 (1H, m), 5.70-5.80 (1H,m), 5.98-6.37 (4H, m), 6.77-7.01 (1H, m), 7.62-7.68 (1H, m), 7.77-7.83(1H, m), 8.03-8.10 (2H, m), 8.14-8.18 (1H, m), 8.41-8.45 (1H, m),8.96-9.01 (1H, m).

ESI-MS m/z 397[M+H]⁺.

Example 27(S)—N-(4-amino-5-(quinolin-3-yl)-8,9-dihydropyrimido[5,4-b]indolizin-8-yl)acrylamide(Compound 27)

Step 1: Synthesis of (S)-tert-butyl(4-amino-5-(quinolin-3-yl)-6,7,8,9-tetrahydropyrimido[5,4-b]indolizin-8-yl)carbamate

A solution of 0.5 M 9-borabicyclo[3.3.1]nonane in tetrahydrofuran (141.3ml) was added to a solution of the (S)-tert-butyl(1-(4-amino-6-bromo-5-(quinolin-3-yl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)but-3-en-2-yl)carbamate(6.0 g) obtained in Step of Example 1 in tetrahydrofuran (42 ml) under anitrogen atmosphere. The mixture was stirred at room temperature for 2hours. A 2 N aqueous sodium hydroxide solution (84.8 ml) was graduallyadded to the reaction mixture at room temperature, and degassed underreduced pressure. Under a nitrogen atmosphere,(tetrakistriphenylphosphine)palladium(0) (1.70 g) was added, and themixture was stirred at 66° C. for 12 hours. After the reaction mixturewas cooled, the organic layer was separated and washed with a 20%aqueous ammonium chloride solution (60 ml). SH silica gel (6.0 g) wasthen added to the organic layer, and the mixture was stirred at 50° C.under a nitrogen atmosphere for 14 hours, and then filtered. SH silicagel (produced by Fuji Silysia Chemical Ltd.) (6.0 g) was added to thefiltrate again, and the mixture was stirred under a nitrogen atmosphereat 50° C. for 14 hours, and then filtered. The solvent was distilled offfrom the filtrate under reduced pressure. The resulting residue waspurified by silica gel column chromatography (developing solvent: ethylacetate/methanol), thereby obtaining the title compound (4.46 g;yield=88%) as a light-yellow solid.

ESI-MS m/z 431 [M+H]⁺.

Step 2: Synthesis of(8S)-4-amino-8-((tert-butoxycarbonyl)amino)-5-(quinolin-3-yl)-6,7,8,9-tetrahydropyrimido[5,4-b]indolizin-6-ylacetate

Tetrabutylammonium iodide (37 mg) and iodobenzene diacetate (241 mg)were added to a solution of the (S)-tert-butyl(4-amino-5-(quinolin-3-yl)-6,7,8,9-tetrahydropyrimido[5,4-b]indolizin-8-yl)carbamate(215 mg) obtained in Step 1 in acetic acid (2 ml) and methylene chloride(2 ml) under ice-cooling. The mixture was stirred for 2 hours underice-cooling and for 2 hours at room temperature. After concentrationunder reduced pressure, the reaction mixture was basified with asaturated aqueous sodium bicarbonate solution, followed by extractionwith ethyl acetate. The resulting organic layer was dried over anhydroussodium sulfate, followed by filtration. Then the solvent was distilledoff under reduced pressure. The resulting residue was purified by silicagel chromatography (developing solvent: ethyl acetate/methanol), therebyobtaining the title compound (216 mg) as a light-brown solid.

ESI-MS m/z 489 [M+H]⁺.

Step 3: Synthesis of tert-butyl((85)-4-amino-6-hydroxy-5-(quinolin-3-yl)-6,7,8,9-tetrahydropyrimido[5,4-b]indolizin-8-yl)carbamate

A 2 N aqueous sodium hydroxide solution (1 ml) was added to a solutionof the(8S)-4-amino-8-((tert-butoxycarbonyl)amino)-5-(quinolin-3-yl)-6,7,8,9-tetrahydropyrimido[5,4-b]indolizin-6-ylacetate (215 mg) obtained in Step 2 in methanol (3 ml) at roomtemperature, and the mixture was stirred for 1 hour at room temperature.The reaction mixture was poured into water, and extracted withchloroform. The resulting organic layer was dried over anhydrous sodiumsulfate, followed by filtration. Then, the solvent was distilled offunder reduced pressure. The resulting residue was purified by silica gelchromatography (developing solvent: ethyl acetate/methanol), therebyobtaining the title compound (166.7 mg) as a light-yellow solid.

ESI-MS m/z 447 [M+H]⁺.

Step 4: Synthesis of(S)-5-(quinolin-3-yl)-8,9-dihydropyrimido[5,4-b]indolizin-4,8-diamine

A mixture of the tert-butyl((8S)-4-amino-6-hydroxy-5-(quinolin-3-yl)-6,7,8,9-tetrahydropyrimido[5,4-b]indolizin-8-yl)carbamate(44.6 mg) obtained in Step 3, p-toluenesulfonic acid monohydrate (95mg), and toluene (3 ml) was stirred for 4 hours at 100° C. After coolingthe reaction mixture, the solvent was distilled off under reducedpressure. The resulting residue was purified by silica gelchromatography (developing solvent: chloroform/methanol), therebyobtaining the title compound (39 mg) as a light-brown, oily substance.

ESI-MS m/z 329 [M+H]⁺.

Step 5: Synthesis of Compound 27

The same synthesis as in Step 10 of Example 13 was performed using the(S)-5-(quinolin-3-yl)-8,9-dihydropyrimido[5,4-b]indolizin-4,8-diamineobtained in Step 4 instead of the(R)-6-methylene-5-(quinolin-3-yl)-6,7,8,9-tetrahydropyrimido[5,4-b]indolizin-4,7-diamineused in Step 10 of Example 13, thereby obtaining the title compound(44.1 mg) as a yellow solid.

¹H-NMR (DMSO-d₆)δ: 4.19-4.30 (2H, m), 4.86-4.94 (1H, m), 5.63 (1H, dd,J=9.8, 2.7 Hz), 6.08-6.32 (5H, m), 6.70 (1H, dd, J=9.8, 1.1 Hz),7.62-7.69 (1H, m), 7.75-7.82 (1H, m), 8.04-8.09 (2H, m), 8.17 (1H, s),8.34 (1H, d, J=2.0 Hz), 8.55 (1H, d, J=7.3 Hz), 8.91 (1H, d, J=2.4 Hz).

ESI-MS m/z 383[M+H]⁺.

Example 28(R)—N-(4-amino-5-(quinolin-3-yl)-9,10-dihydro-8H-pyrimido[5′,4′:4,5]pyrrolo[1,2-a]azepin-8-yl)acrylamide(Compound 28)

The same synthesis as in Example 27 was performed using the(R)-tert-butyl(5-(4-amino-6-bromo-5-(quinolin-3-yl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)pent-1-en-3-yl)carbamateobtained in Step 7 of Example 13 instead of the (S)-tert-butyl(1-(4-amino-6-bromo-5-(quinolin-3-yl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)but-3-en-2-yl)carbamateused in Step 1 of Example 27, thereby obtaining the title compound(101.8 mg) as a yellow solid.

¹H-NMR (DMSO-d₆)δ: 2.01-2.12 (1H, m), 2.23-2.33 (1H, m), 4.28-4.38 (1H,m), 4.55-4.63 (1H, m), 4.84-4.92 (1H, m), 5.62 (1H, dd, J=10.0, 2.4 Hz),5.68 (1H, dd, J=12.6, 3.8 Hz), 6.08-6.31 (5H, m), 7.63-7.69 (1H, m),7.78-7.83 (1H, m), 8.03-8.10 (2H, m), 8.18 (1H, s), 8.31 (1H, d, J=2.0Hz), 8.51 (1H, d, J=8.2 Hz), 8.84 (1H, d, J=2.2 Hz).

ESI-MS m/z 397[M+H]⁺.

Example 29

N-((6R*,8S)-4-amino-6-methyl-5-(quinolin-3-yl)-6,7,8,9-tetrahydropyrimido[5,4-b]indolizin-8-yl)acrylamide(Compound 29A) andN-H6S*,85)-4-amino-6-methyl-5-(quinolin-3-yl)-6,7,8,9-tetrahydropyrimido[5,4-b]indolizin-8-yl)acrylamide(Compound 29B)

Step 1: Synthesis of (S)-tert-butyl(4-amino-6-methylene-5-(quinolin-3-yl)-6,7,8,9-tetrahydropyrimido[5,4-b]indolizin-8-yl)carbamate

A mixture of the (S)-tert-butyl(4-amino-6-methyl-5-(quinolin-3-yl)-8,9-dihydropyrimido[5,4-b]indolizin-8-yl)carbamateand (S)-tert-butyl(4-amino-6-methylene-5-(quinolin-3-yl)-6,7,8,9-tetrahydropyrimido[5,4-b]indolizin-8-yl)carbamateobtained by the same synthesis as in Step 5 of Example 14 using the(S)-tert-butyl(1-(4-amino-6-bromo-5-(quinolin-3-yl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)pent-4-en-2-yl)carbamate(1.263 g) obtained in Step 4 of Example 14 was purified by silica gelchromatography (developing solvent: ethyl acetate/methanol), therebyobtaining the title compound (0.805 g) as a light-yellow solid.

ESI-MS m/z 443 [M+H]⁺.

Step 2: Synthesis of tert-butyl((85)-4-amino-6-methyl-5-(quinolin-3-yl)-6,7,8,9-tetrahydropyrimido[5,4-b]indolizin-8-yl)carbamate

10% palladium-carbon (50% wet, 100.6 mg) was added to a solution of the(S)-tert-butyl(4-amino-6-methylene-5-(quinolin-3-yl)-6,7,8,9-tetrahydropyrimido[5,4-b]indolizin-8-yl)carbamate(100.0 mg) obtained in Step 1 in methanol (10 ml), and the mixture wasstirred for 12 hours at room temperature under a hydrogen atmosphere.The reaction mixture was filtered through a glass fiber filter, and thefiltrate was concentrated under reduced pressure. The resulting residuewas purified by basic silica gel column chromatography (developingsolvent: chloroform/methanol), thereby obtaining the title compound(122.1 mg).

ESI-MS m/z 445 [M+H]⁺.

Step 3: Synthesis of(85)-6-methyl-5-(quinolin-3-yl)-6,7,8,9-tetrahydropyrimido[5,4-b]indolizin-4,8-diamine

Trifluoroacetic acid (1 ml) was added to a solution of the tert-butyl((8S)-4-amino-6-methyl-5-(quinolin-3-yl)-6,7,8,9-tetrahydropyrimido[5,4-b]indolizin-8-yl)carbamate(122.1 mg) obtained in Step 2 in chloroform (3 ml), and the mixture wasstirred for 30 minutes at room temperature. The reaction mixture wasconcentrated under reduced pressure, and the resulting residue wasdissolved in methanol. After desalting with a basic solid-phaseextraction column (VARIAN BondElut), the mixture was concentrated underreduced pressure, thereby obtaining the title compound (58.9 mg) as alight-yellow solid.

ESI-MS m/z 345 [M+H]⁺.

Step 4: Synthesis ofN-H8S)-4-amino-6-methyl-5-(quinolin-3-yl)-6,7,8,9-tetrahydropyrimido[5,4-b]indolizin-8-yl)acrylamide

The same synthesis as in Step 7 of Example 14 was performed using the(8S)-6-methyl-5-(quinolin-3-yl)-6,7,8,9-tetrahydropyrimido[5,4-b]indolizin-4,8-diamine(58.9 mg) obtained in Step 3, thereby obtaining the title compound (30.3mg) as a light-yellow solid.

ESI-MS m/z 399 [M+H]⁺.

Step 5: Syntheses of Compound 29A and Compound 29B

A diastereomer mixture containing the (6R*,8S) isomer and (6S*,8S)isomer obtained in Step 4 was separated by a preparative chiral columnchromatography (column: CHIRALCEL OZ-H (20 mm×250 mm×5 μm), manufacturedby Daicel Chemical Industries, Ltd., developing solvent:hexane/ethanol/triethylamine=60/40/0.01), thereby obtaining Compound 29Aand Compound 29B as a first fraction (15.8 mg) and a second fraction(6.1 mg), respectively, as colorless solids.

Compound 29A

¹H-NMR (CDCl₃)δ: 0.96 (3H, d, J=6.6 Hz), 1.68 (1H, m), 2.29-2.39 (1H,m), 3.49-3.63 (1H, m), 3.81 (1H, dd, J=11.7, 10.2 Hz), 4.56-4.69 (1H,m), 4.76 (1H, dd, J=12.2, 5.1 Hz), 4.81 (2H, brs), 5.70 (1H, dd, J=10.5,1.2 Hz), 6.17 (1H, dd, J=16.6, 10.2 Hz), 6.15-6.25 (1H, m), 6.36 (1H,dd, J=17.1, 1.2 Hz), 7.64 (1H, t, J=7.6 Hz), 7.75-7.82 (1H, m), 7.88(1H, d, J=8.0 Hz), 8.18 (1H, d, J=8.5 Hz), 8.21-8.29 (2H, m), 9.05 (1H,d, J=2.0 Hz).

ESI-MS m/z 399[M+H]⁺.

Compound 29B

¹H-NMR (CDCl₃)δ: 1.06 (3H, d, J=6.8 Hz), 1.83-2.08 (1H, m), 2.18-2.31(1H, m), 3.46-3.59 (1H, m), 4.00 (1H, dd, J=12.4, 7.6 Hz), 4.59 (1H, dd,J=12.4, 4.6 Hz), 4.73-4.93 (3H, m), 5.70 (1H, d, J=10.5 Hz), 6.18 (1H,dd, J=16.8, 10.2 Hz), 6.26-6.42 (2H, m), 7.63 (1H, t, J=7.6 Hz), 7.78(1H, t, J=8.3 Hz), 7.88 (1H, d, J=8.0 Hz), 8.17 (1H, d, J=8.0 Hz),8.20-8.27 (2H, m), 9.02 (1H, d, J=1.7 Hz).

ESI-MS m/z 399[M+H]⁺.

Comparative Example 1N-(3-(4-amino-6,7,8,9-tetrahydropyrimido[5,4-b]indolizin-5-yl)phenyl)benzamide(Comparative Compound 1)

The compound was synthesized according to the method disclosed in theInternational Publication No. WO2006/102079 pamphlet.

ESI-MS m/z 384 [M+H]⁺.

Test Examples

The compounds of the present invention were evaluated using the testmethods below.

Test Example 1 Measurement of Inhibitory Action Against Various EGFRKinase Activities (In Vitro)

1) Measurement of EGFR (T790M/L858R) Kinase Inhibitory Activity

The inhibitory activities of the compounds of the present inventionagainst EGFR (T790M/L858R) kinase activity were measured.

Among the materials of the inhibitory activity measurement, thesubstrate peptide and the kinase protein were obtained as follows. Apeptide with biotinylated N-terminus (biotin-EEPLYWSFPAKKK) wassynthesized with reference to FL-Peptide 22, a reagent for LabChip®series of PerkinElmer, Inc. For the kinase protein, a purifiedrecombinant human EGFR (T790M/L858R) protein of Carna Biosciences, Inc.was purchased.

The inhibitory activity measurement was performed as follows. Thecompounds of the present invention were individually dissolved indimethyl sulfoxide (DMSO), and serial dilutions of these compounds wereprepared using DMSO. Subsequently, a solution containing the substratepeptide (final concentration: 250 nM), magnesium chloride (finalconcentration: 10 mM), manganese chloride (final concentration: 10 mM),and ATP (final concentration: 1 μM) in a buffer solution for kinasereaction (Carna Biosciences, Inc.) was mixed with the dilutions of thecompounds (final concentration of DMSO upon kinase reaction: 2.5%) orDMSO (final concentration: 2.5%). EGFR (T790M/L858R) protein was furtheradded thereto, and the mixtures were incubated at 25° C. for 120 minutesto carry out a kinase reaction. EDTA was added thereto to a finalconcentration of 24 mM to thereby terminate the reaction. Aphosphorylated thyrosine detection solution containing europium(Eu)-labeled anti-phosphorylated tyrosine antibody PT66 (PerkinElmer,Inc.) and SureLight APC-SA (PerkinElmer, Inc.) was added to the reactionmixtures, and the mixtures were allowed to stand at room temperature for2 hours or more. For the background, a sample to which EDTA was addedbefore addition of EGFR (T790M/L858R) protein was incubated at 25° C.for 120 minutes, using DMSO instead of the DMSO solution of thecompound. The detection solution was also added to this sample, and themixture was allowed to stand for 2 hours or more. Finally, for all ofthe test samples, the amount of fluorescence at the time of irradiationof excitation light having a wavelength of 337 nm was measured at dualwavelengths of 620 nm and 665 nm by PHERAstar FS (BMG LABTECH), and aratio of the fluorescence amounts at the two wavelengths was obtained asdata.

In the analysis of measured data, a ratio of the fluorescence amounts ofthe sample in which the kinase reaction was carried out with theaddition of DMSO at a final concentration of 2.5% was set as aphosphorylation inhibition rate of 0%, and a ratio of the fluorescenceamounts of the background was set as a phosphorylation inhibition rateof 100%. Based on the above rates, phosphorylation inhibition rates (%)were calculated for the samples to which various concentrations of thecompound solutions were added. Finally, the obtained reaction inhibitionrates (%) at the respective concentrations were plotted for eachcompound, and the IC₅₀ value (nM), which is a compound concentration atwhich phosphorylation by EGFR (T790M/L858R) is inhibited by 50%, wasdetermined using XLfit curve-fitting software (IDBS).

2) Measurement of EGFR (d746-750/T790M) Kinase Inhibitory Activity

The inhibitory activities of the compounds of the present inventionagainst EGFR (d746-750/T790M) kinase activity were measured.

The materials, the measurement method, and the data analysis method weresubstantially the same as those shown in the above description regarding“Measurement of EGFR (T790M/L858R) Kinase Inhibitory Activity,” exceptthat a purified recombinant human EGFR (d746-750/T790M) proteinpurchased from Carna Biosciences, Inc. was used as the kinase protein,and the measurement was performed with an ATP final concentration of 1.5μM. Finally, the IC₅₀ value (nM) of each compound with respect to EGFR(d746-750/T790M) was determined by data analysis.

3) Measurement of EGFR (L858R) Kinase Inhibitory Activity

The inhibitory activities of the compounds of the present inventionagainst EGFR (L858R) kinase activity were measured.

The materials, the measurement method, and the data analysis method weresubstantially the same as those shown in the above description regarding“Measurement of EGFR (T790M/L858R) Kinase Inhibitory Activity,” exceptthat a purified recombinant human EGFR (L858R) protein purchased fromCarna Biosciences, Inc. was used as the kinase protein, and themeasurement was performed with ATP at a final concentration of 4 μM.Finally, the IC₅₀ value (nM) of each compound with respect to EGFR(L858R) was determined by data analysis.

4) Measurement of EGFR (d746-750) Kinase Inhibitory Activity

The inhibitory activities of the compounds of the present inventionagainst EGFR (d746-750) kinase activity were measured.

The materials, the measurement method, and the data analysis method weresubstantially the same as those shown in the above description regarding“Measurement of EGFR (T790M/L858R) Kinase Inhibitory Activity,” exceptthat a purified recombinant human EGFR (d746-750) protein purchased fromCarna Biosciences, Inc. was used as the kinase protein, the measurementwas performed with ATP at a final concentration of 5 μM, and theincubation for the kinase reaction was performed for 90 minutes.Finally, the IC₅₀ value (nM) of each compound with respect to EGFR(d746-750) was determined by data analysis.

5) Measurement of EGFR (WT) Kinase Inhibitory Activity

The inhibitory activities of the compounds of the present inventionagainst EGFR (WT) kinase activity were measured.

The materials, the measurement method, and the data analysis method weresubstantially the same as those shown in the above description regarding“Measurement of EGFR (T790M/L858R) Kinase Inhibitory Activity,” exceptthat a kinase protein prepared by expressing a cytoplasmic domain ofhuman EGFR (WT) with its N-terminus fused to a FLAG tag in insect Sf9cells using a baculovirus expression system and then purifying it withanti-FLAG antibody agarose (Sigma-Aldrich) was used as the kinaseprotein, the final concentration of the substrate peptide was 500 nM,and the final concentration of ATP was 4.7 pM. Finally, the IC₅₀ value(nM) of each compound with respect to EGFR (WT) was determined by dataanalysis.

Table 1 shows the results of the tests using the various EGFR.

It was confirmed that the compounds of the present invention had stronginhibitory activities not only for EGFR (L858R) and EGFR (d746-750), butalso for EGFR (T790M/L858R) and EGFR (d746-750/T790M). It was alsoconfirmed that their inhibitory activities for EGFR (WT) were lower thanthose for the above EGFR proteins.

In contrast, it was confirmed that the compound of Comparative Example1, which was a compound having a structure similar to that of thecompound of the present invention, had almost no inhibitory activityagainst these EGFR kinases.

TABLE 1 Type of EGFR EGFR EGFR EGFR (T790M/ (d746-750/ EGFR (d746- EGFRExample L858R) T790M) (L858R) 750) (WT) 1 <0.50 <0.15 <0.50 <0.15 0.70 277 30 230 78 1400 3 6.1 1.8 15 7.6 110 4 0.40 <0.50 0.63 0.45 6.0 5<0.15 <0.15 <0.15 <0.15 0.78 6 0.24 <0.15 0.45 0.24 4.9 7 1.0 0.53 1.71.2 12 8 <0.50 <0.15 <0.50 <0.15 1.0 9 2.1 0.45 5.9 2.3 52 10 1.5 0.612.6 1.8 22 11 0.76 0.32 1.4 0.88 12 12 0.66 0.29 1.2 0.75 11 13 2.1 1.12.8 2.1 24 14 0.24 <0.50 0.32 0.19 2.9 15 1.6 1.2 1.3 1.4 7.9 16 11 3.039 15 480 17 23 11 50 39 580 18 4.7 1.3 12 4.5 68 19 0.26 0.28 0.27 0.422.2 20 3.7 2.4 4.1 2.9 28 21 0.60 0.34 1.1 0.72 7.8 22 0.35 0.23 0.370.33 5.5 23 0.44 0.30 0.63 0.29 7.3 24 0.83 0.31 1.1 0.85 8.6 25 36 11100 59 26 1.0 0.31 1.7 1.1 27 0.34 0.25 0.35 0.38 28 0.30 <0.50 0.490.27 29 14 8.4 19 21 Comparative >5000 >5000 >5000 1500 >5000 Example 1

Test Example 2 Test for Activity of Growth Inhibition against Wild-Typeand Mutated EGFR-Expressing Cell Lines (In Vitro)

(1) A human small cell lung cancer cell line NCI-H1975 expressing EGFR(T790M/L858R), (2) a human non-small cell lung cancer cell line HCC827expressing EGFR (d746-750), and (3) a human epithelioid cancer cell lineA431 expressing EGFR (WT) were each suspended in complete growth mediumrecommended by ATCC. The cell suspensions were seeded in each well of a384-well flat microplate or a 96-well flat plate, and cultured in anincubator containing 5% carbon dioxide gas at 37° C. for one day. Eachcompound of the present invention was dissolved in DMSO, and serialdilutions of the test compound were prepared using DMSO (to aconcentration 500 times the final concentration). The DMSO solution ofthe test compound or DMSO was diluted with complete growth medium, andthe resulting solution was added to each well of the cell culture plateso that the final concentration of DMSO was 0.2%. Then, the cells werecultured in an incubator containing 5% carbon dioxide gas at 37° C. forthree days. The number of cells was measured before the addition of theDMSO solution of the test compound, as well as after addition andculture by using a CellTiter-Glo Assay® (produced by Promega) accordingto a protocol recommended by Promega.

For each cell, the cell growth inhibition rates of the wells to whichthe test compounds at various concentrations were added were calculatedby the following formula. Then, the obtained inhibition rates (%) at therespective concentrations were plotted for each test compound, and theGI₅₀ value (nM), which is a test compound concentration at which thecell growth can be inhibited by 50%, was determined using XLfitcurve-fitting software (IDBS).

-   Cell Growth Inhibition Rate (%)=(C−T)/(C−CO)×100-   T: Luminescence intensity of well cultured for three days after    addition of the test compound solution-   C: Luminescence intensity of well cultured for three days after    addition of DMSO-   C0: Luminescence intensity of well before addition of the test    compound solution

Table 2 shows the results.

It was confirmed that the compounds of the present invention showedstrong growth inhibitory effects not only for EGFR (d746-750) expressingcells, but also for EGFR (T790M/L858R) expressing cells. It was alsoconfirmed that their growth inhibitory effects for EGFR (WT) expressingcells were lower than those for the above EGFR proteins.

TABLE 2 Type of EGFR EGFR EGFR EGFR (T790M/L858R) (d746-750) (WT) Cellname Example NCI-H1975 HCC827 A431 1 9.0 1.9 370 2 4200 780 >10000 3 56016 >10000 4 140 5.5 2400 5 51 8.1 2200 6 54 3.3 520 7 150 5.6 2600 8 80066 1100 9 770 7.5 4200 10 310 8.1 4400 11 260 9.7 3900 12 240 7.9 350013 150 13 1200 14 22 2.5 1900 15 85 22 2000 16 600 27 >10000 17 2900160 >10000 18 160 8.9 4000 19 49 5.6 990 20 310 23 3900 21 56 8.1 260022 56 5.7 1500 23 47 3.3 5100 24 110 4.9 3400 25 2500 260 >10000 26 15018.4 4700 27 110 15 1000 28 48 14 1200 29 1600 89 >10000

The invention claimed is:
 1. A compound represented by Formula (I) below or a salt thereof:

wherein the group:

is a group represented by Formula A3:

wherein in Formula A3, B is a group represented by:

wherein R₁ is a hydrogen atom or a C1-C6 alkyl group; and R₂ is a group represented by:

wherein R₃, R₄, and R₅ are the same or different, and each represents a hydrogen atom, a halogen atom, a C1-C6 alkyl group, a C6-C12 aryl group, a C4-C9 heteroaryl group, an aminomethyl group that may be substituted with a Cl -C6 alkyl group, or a 1-piperidinomethyl group, or a group represented by:

wherein R₆ represents a hydrogen atom or a C1-C6 alkyl group, m is 0 or 1; and n is 1 or 2; and R₁₀ is methyl.
 2. The compound or a salt thereof according to claim 1, wherein R₂ is a group represented by:

wherein R₃, R₄, and R₅ are the same or different, and each represents a hydrogen atom, a halogen atom, a C1-C6 alkyl group, an aminomethyl group that may be substituted with a C1-C6 alkyl group, or a 1-piperidinomethyl group, or a group represented by:

wherein R₆ represents a hydrogen atom or a C1-C6 alkyl group.
 3. The compound or a salt thereof according to claim 1, wherein R₂ is a group represented by:

wherein R₃, R₄, and R₅ are the same or different, and each represents a hydrogen atom, a halogen atom, an aminomethyl group that may be substituted with a methyl group, or a 1-piperidinomethyl group.
 4. The compound or a salt thereof according to claim 1, wherein the compound is selected from the following group of compounds: N-((7S)-4-amino-6-methyl-5-(quinolin-3-yl)-7,8-dihydro-6H-pyrimido [5 ,4-b]pyrrolizin-7-yl)acrylamide; N-((6R*,8S)-4-amino-6-methyl-5-(quinolin-3-yl)-6,7,8,9-tetrahydropyrimido[5,4-b]indolizin-8-yl)acrylamide, and N-((6S *,8S)-4-amino-6-methyl-5-(quinolin-3-yl)-6,7,8,9-tetrahydropyrimido [5 ,4-b] indolizin-8-yl)acrylamide.
 5. An EGFR inhibitor comprising the compound or a salt thereof according to claim 1, as an active ingredient.
 6. A pharmaceutical composition comprising the compound or a salt thereof according to claim
 1. 7. An antitumor agent comprising the compound or a salt thereof according to claim 1, as an active ingredient.
 8. A method for inhibiting EGFR in a mammal in need thereof, comprising administering, to the mammal, a compound or a salt thereof according to claim 1 at a dose effective for inhibiting EGFR, wherein said mammal has cancer.
 9. A method for the manufacture of an antitumor agent, comprising combining the compound or a salt thereof according to claim 1 with a pharmaceutical carrier acceptable for use in an antitumor agent.
 10. The compound or a salt thereof according to claim 1 for use in the prevention or treatment of cancer. 